Compositions of S-nitrosothiols and methods of use

ABSTRACT

The invention describes compositions and kits comprising 4-aza-4-(2-methyl-2-sulfanylpropyl)tricyclo(5.2.1.0&lt;2,6&gt;)dec-8-ene-3,5-dione; 4-aza-4-(2-methyl-2-(nitrosothio)propyl)tricyclo(5.2.1.0&lt;2,6&gt;)dec-8-ene-3,5-dione; 4-{1-methyl-1-((2,4,6-trimethoxyphenyl)methylthio)ethyl}-1,3-oxazolidin-2-one; 2-aAmino-3-methyl-3-((2,4,6-trimethoxyphenyl)methylthio)butan-1-ol; 4-(1-methyl- 1 -(nitrosothio)ethyl)-1,3-oxazolidin-2-one; or pharmaceutically acceptable salts thereof. The compositions of the invention can further comprise at least one penetration enhancer, at least one compound that donates, transfers or releases nitric oxide, elevates endogenous levels of endothelium-derived relaxing factor, stimulates endogenous synthesis of nitric oxide or is a substrate for nitric oxide synthase and/or at least one vasoactive agent.

RELATED APPLICATIONS

[0001] This application is a continuation under 35 USC § 120 of U.S.application Ser. No. 09/850,081 filed May 8, 2001, now allowed, whichclaims priority under 35 USC § 119 to U.S. Application No. No.60/202,935 filed May 9, 2000.

FIELD OF THE INVENTION

[0002] The present invention describes rapid noninvasive methods formeasuring vasodilation or changes in blood flow in a patient followingadministration of at least one compound that donates, transfers orreleases nitric oxide, elevates endogenous levels of endothelium-derivedrelaxing factor, stimulates endogenous synthesis of nitric oxide or is asubstrate for nitric oxide synthase and/or at least one vasoactiveagent. The method comprises the administration of at least one compoundthat donates, transfers or releases nitric oxide, elevates endogenouslevels of endothelium-derived relaxing factor, stimulates endogenoussynthesis of nitric oxide or is a substrate for nitric oxide synthaseand/or at least one vasoactive agent to the patient followed bymonitoring the temperature change of an area of interest using infraredthermography. The present invention provides methods for diagnosingdiseases or disorders related to vasodilation and changes in blood flow,such as, sexual dysfunctions, Raynaud's syndrome, inflammation,hypertension, gastrointestinal disorders and central nervous systemdisorders. The sexual dysfunction is preferably female sexualdysfunction or female sexual arousal. The vasoactive agents includepotassium channel activators, calcium channel blockers, α-adrenergicreceptor antagonists, β-blockers, phosphodiesterase inhibitors,adenosine, ergot alkaloids, vasoactive intestinal peptides,prostaglandins, dopamine agonists, opioid antagonists, endothelinantagonists and thromboxane inhibitors. The present invention can alsobe used to screen and identify drug candidates for treating diseases,disorders and conditions resulting from vasodilation or changes in bloodflow. The present invention also describes compositions comprising atleast one S-nitrosothiol compound for diagnosing, monitoring and/ortreating female sexual dysfunctions.

BACKGROUND OF THE INVENTION

[0003] Adequate sexual function is a complex interaction of hormonalevents and psychosocial relationships. There are four stages to sexualresponse as described in the International Journal of Gynecology &Obstetrics, 51(3):265-277 (1995). The first stage of sexual response isdesire. The second stage of sexual response is arousal. Both physicaland emotional stimulation may lead to breast and genital vasodilationand clitoral engorgement (vasocongestion). In the female, dilation andengorgement of the blood vessels in the labia and tissue surrounding thevagina produce the “orgasmic platform,” an area at the distal third ofthe vagina where blood becomes sequestered. Localized perivaginalswelling and vaginal lubrication make up the changes in this stage ofsexual response. Subsequently, ballooning of the proximal portion of thevagina and elevation of the uterus occurs. In the male, vasodilation ofthe cavernosal arteries and closure of the venous channels that drainthe penis produce an erection. The third stage of sexual response isorgasm, while the fourth stage is resolution. Interruption or absence ofany of the stages of the sexual response cycle can result in sexualdysfunction. One study found that 35% of males and 42% of femalesreported some form of sexual dysfunction. Read et al, J. Public HealthMed., 19(4):387-391 (1997).

[0004] While there are obvious differences in the sexual responsebetween males and females, one common aspect of the sexual response isthe erectile response. The erectile response in both males and femalesis the result of engorgement of the erectile tissues of the genitaliawith blood which is caused by the relaxation of smooth muscles in thearteries serving the genitalia. This increase in blood flow results invasodilation and an increase in the temperature of the genitalia tissue.

[0005] Methods for evaluating and measuring physiological changes todetermine sexual arousal have been previously described. For example,U.S. Pat. Nos. 5,565,466 and 5,731,339 describe the use of Dopplerultrasound velocimetry for measuring vaginal and penile blood flow;Intl. J. Impotence Res., 9:27-37 (1997) discloses the use ofphotoplethysmorgraphy for monitoring vaginal blood flow; and WO 99/35968describes devices and methods for monitoring female arousal. These priorart methods are invasive and have major limitations that can effect theaccuracy of the measurements.

[0006] There is a need in the art for new and improved noninvasivemethods for measuring vasodilation and changes in blood flow, and fordiagnosing and monitoring diseases related to vasodilation, such as,sexual dysfunctions. The present invention is directed to these, as wellas other, important ends.

SUMMARY OF THE INVENTION

[0007] In arriving at the present invention, it was unexpectedlydiscovered that the vasodilation or engorgement of the corpus cavernosumsmooth muscle, an event involved in the sexual response process in bothmales and females, results in an increase in temperature that can bemonitored by infrared thermography.

[0008] One aspect of the present invention describes methods formonitoring and measuring vasodilation and changes in blood flow inpatients, by administering to a patient in need thereof at least onecompound that donates, transfers or releases nitric oxide, elevatesendogenous levels of endothelium-derived relaxing factor, stimulatesendogenous synthesis of nitric oxide or is a substrate for nitric oxidesynthase and/or at least one vasoactive agent, followed by monitoringthe temperature change of an area of interest using infraredthermography. Another aspect of the invention comprises the steps ofcomparing the temperature of the area prior to and after theadministration of the nitric oxide donor and/or vasoactive agent. Thusthe method may comprise (i) measuring the baseline temperature of anarea of interest before exposure to at least one nitric oxide donorand/or vasoactive agent using infrared thermography, (ii) administeringto a patient at least one nitric oxide donor and/or vasoactive agent,(iii) measuring the temperature of the area of interest during and/orafter administering of at least one nitric oxide donor and/or vasoactiveagent using infrared thermography, and (iv) comparing the measurementsobtained in steps (i) and (iii), wherein a compound that results in anincrease in the temperature is a vasodilator and a compound that resultsin a decrease in the temperature is a vasoconstrictor. The temperaturein step (i) may be a previously obtained stable temperature measurement;or alternatively a measurement taken any time after the administrationof the nitric oxide donor and/or vasoactive agent when a stabletemperature measurement is obtained. The nitric oxide donors, and/orvasoactive agents can be administered separately or as components of thesame composition in one or more pharmaceutically acceptable carriers.

[0009] Another aspect of the invention provides methods for monitoringand diagnosing diseases and disorders related to blood flow, such as,sexual dysfunctions and sexual arousal in patients, preferably males andfemales, more preferably females, by administering to a patient in needthereof at least one compound that donates, transfers or releases nitricoxide, elevates endogenous levels of endothelium-derived relaxingfactor, stimulates endogenous synthesis of nitric oxide or is asubstrate for nitric oxide synthase and/or at least one vasoactiveagent, followed by measuring the temperature of the genitalia usinginfrared thermography or followed by comparing the temperature of thegenitalia after administration of the at least one nitric oxide donorand/or vasoactive agent with a stable baseline temperature measurementobtained using infrared thermography.

[0010] Yet another aspect of the invention provides methods formonitoring and diagnosing diseases and disorders resulting fromvasodilation and changes in blood flow by administering to a patient inneed thereof at least one compound that donates, transfers or releasesnitric oxide, elevates endogenous levels of endothelium-derived relaxingfactor, stimulates endogenous synthesis of nitric oxide or is asubstrate for nitric oxide synthase and/or at least one vasoactiveagent, followed by comparing the temperature change of the area ofinterest after administration of the at least one nitric oxide donorand/or vasoactive agent with a stable baseline temperature measurementusing infrared thermography. The diseases and disorders resulting fromchanges in blood flow include Raynaud's syndrome, inflammation,hypertension, gastrointestinal disorders and central nervous systemdisorders.

[0011] Another aspect of the present invention comprises identification,characterization, rank and selection of compounds that are can be usedto treat numerous diseases and disorders resulting from vasodilation andchanges in blood flow. For example a test compound may be administeredto a patient followed by infrared thermographic measurements of an areaof interest to monitor the temperature changes. Compounds that result ina temperature increase are effective for the treatment of the disease ordisorder, the greater the increase in temperature, the more potent thecompound as a suitable therapy. On the other hand, compounds thatproduce no temperature change or that result in a temperature decreaseare not effective for the treatment of the disease or disorder, thegreater the decrease in temperature, the less effective the compound asa suitable therapy.

[0012] Yet another aspect of the present invention provides compositionscomprising at least one S-nitrosothiol compound and at least onepenetration enhancer that may be used to diagnose, monitor and/or treatfemale sexual dysfunctions. The S-nitrosothiol compound may preferablybe S-nitrosoglutathione. The penetration enhancer, may preferably be aglyceride, such as, MIGLYOL® and/or a polyglycolyzed glyceride, such as,LABROSOL® and/or LABROFIL®, or a mixture thereof. These compositions mayfurther comprise at least one vascoactive agent and/or at least onenitric oxide donor, or mixtures thereof.

[0013] These and other aspects of the present invention are described indetail herein.

BRIEF DESCRIPTION OF THE FIGURES

[0014]FIG. 1 shows the infrared thermographic images following topicaladministration of 10% isobutryl nitrate (top panels) or vehicle (bottompanels) to a rabbit vagina and clitoris. The x axis corresponds to timein minutes from just prior to application of isobutryl nitrate orvehicle (0 minutes) to 30 minutes after application of isobutryl nitrateor vehicle. The vertical bar on the left hand side corresponds to thecolor change for temperatures ranging from 28° C. to 36° C.

[0015]FIG. 2 shows the infrared thermographic images following topicaladministration of 20% isobutryl nitrate (top panels) or vehicle (bottompanels) to a rabbit vagina and clitoris. The x axis corresponds to timein minutes from just prior to application of isobutryl nitrate orvehicle (0 minutes) to 30 minutes after application of isobutryl nitrateor vehicle. The vertical bar on the left hand side corresponds to thecolor change for temperatures ranging from 26° C. to 36° C.

[0016]FIG. 3 shows the infrared thermographic images following topicaladministration of Example 1 (top panels) or vehicle (bottom panels) to arabbit vagina and clitoris. The x axis corresponds to time in minutesfrom just prior to application of Example 1 or vehicle (0 minutes) to 60minutes after application of Example 1 or vehicle. The vertical bar onthe left hand side corresponds to the color change for temperaturesranging from 30° C. to 36° C.

[0017]FIG. 4 shows the infrared thermographic images following topicaladministration of 10% Example 2 (top panels) to rabbit vagina andclitoris. The x axis corresponds to time in minutes from just prior toapplication of Example 2 (0 minutes) to 60 minutes after application ofExample 2. The bottom panels show the effect of administration of 10%phenylephrine (PE 10%, first bottom panel). The x axis corresponds totime in minutes for 5 minutes after the application of phenylephrinefrom 61 minutes to 65 minutes. The vertical bar on the left hand sidecorresponds to the color change for temperatures ranging from 26° C. to38° C.

[0018]FIG. 5 shows the change in temperature, as measured by infraredthermography, following the topical administration of (a) Example 7(10%, closed circles); (b) Example 7 (5%, closed squares); (c) Example 7(2.5%, closed diamonds); or (d) vehicle alone (MIGLYOL, closedtrianges); to a rabbit vagina and clitoris. The x axis corresponds totime in minutes. The y axis corresponds to the change in temperature (°C) after the topical administration of the test compound.

[0019]FIG. 6 shows the change in blood flow, as measured by a laserdoppler probe, following the topical administration of (a) Example 7(10%, closed circles); (b) Example 7 (5%, closed squares); or (c)vehicle alone (MIGLYOL®, closed triangles); to a rabbit vagina andclitoris. The x axis corresponds to time in minutes. The y axiscorresponds to the change in temperature (° C.) after the topicaladministration of the test compound.

DETAILED DESCRIPTION OF THE INVENTION

[0020] As used throughout the disclosure, the following terms, unlessotherwise indicated, shall be understood to have the following meanings.

[0021] “Patient” refers to animals, preferably mammals, more preferablyhumans, and includes children and adults, and males and females.

[0022] “Infrared thermography” refers to the recording of thetemperature of a body by means of infrared radiation emitted by thesurface of the body at wavelengths of between about 0.8 μm and about 1mm. The monitoring of radiation is preferably in the range of about 3 μmto about 100 μm, more preferably in the range of about 3 μm to about 15μm, and most preferably in the range of about 3 μm to about 12 μm.

[0023] “Area of interest” refers to the area whose temperature isrecorded and monitored using infrared thermography. The area of interestmay include the symptomatic area.

[0024] “Baseline temperature” refers to the temperature of the area ofinterest at rest i.e., without the administration of a compound. Thebaseline temperature can be measured at, for example, prior to theadministration of the test compound i.e., nitric oxide donor and/orvasoactive agent. Alternatively, the baseline temperature can bemeasured after the administration of the nitric oxide donor and/orvasoactive agent when a stable temperature reading is obtained.

[0025] “Vasoactive agent” refers to any therapeutic agent capable ofrelaxing vascular and/or nonvascular smooth muscle. Suitable vasoactiveagents include, but are not limited to, potassium channel activators,calcium channel blockers, β-blockers, long and short acting α-adrenergicreceptor antagonists, prostaglandins, phosphodiesterase inhibitors,adenosine, ergot alkaloids, vasoactive intestinal peptides, dopamineagonists, opioid antagonists, endothelin antagonists, thromboxaneinhibitors, and the like.

[0026] “Thromboxane inhibitor” refers to any compound that reversibly orirreversibly inhibits thromboxane synthesis, and includes compoundswhich are the so-called thromboxane A₂ receptor antagonists, thromboxaneA₂ antagonists, thromboxane A₂/prostaglandin endoperoxide antagonists,thromboxane receptor (TP) antagonists, thromboxane antagonists,thromboxane synthase inhibitors, and dual acting thromboxane synthaseinhibitors and thromboxane receptor antagonists.

[0027] “Thromboxane A₂ receptor antagonist” refers to any compound thatreversibly or irreversibly blocks the activation of any thromboxane A₂receptor.

[0028] “Thromboxane synthase inhibitor” refers to any compound thatreversibly or irreversibly inhibits the enzyme thromboxane synthesisthereby reducing the formation of thromboxane A₂.

[0029] “Dual acting thromboxane receptor antagonist and thromboxanesynthase inhibitor” refers to any compound that simultaneously acts as athromboxane A₂ receptor antagonist and a thromboxane synthase inhibitor.

[0030] “Raynaud's syndrome” refers to a condition that causes a loss ofblood flow to the fingers, toes, nose and/or ears. The affected areaturns white from the lack of circulation, then blue and cold, andfinally numb. The affected area may also turn red, and may throb, tingleor swell.

[0031] “Gastrointestinal disorder” refers to any disease or disorder ofthe upper gastrointestinal tract of a patient including, for example,inflammatory bowel disease, peptic ulcers, stress ulcers, gastrichyperacidity, dyspepsia, gastroparesis, Zollinger-Ellison syndrome,gastroesophageal reflux disease, short-bowel (anastomosis) syndrome,hypersecretory states associated with systemic mastocytosis orbasophilic leukemia and hyperhistaminemia, and bleeding peptic ulcersthat result, for example, from neurosurgery, head injury, severe bodytrauma or burns.

[0032] “Upper gastrointestinal tract” refers to the esophagus, thestomach, the duodenum and the jejunum.

[0033] “Ulcers” refers to lesions of the upper gastrointestinal tractlining that are characterized by loss of tissue. Such ulcers includegastric ulcers, duodenal ulcers and gastritis.

[0034] “Therapeutically effective amount” refers to the amount of thecompound and/or composition that is effective to achieve its intendedpurpose.

[0035] “Topical” refers to the delivery of a compound by passage throughthe skin and into the blood stream and includes transdermal delivery.

[0036] “Transdermal” refers to the delivery of a compound by passagethrough the skin and into the blood stream.

[0037] “Transmucosal” refers to delivery of a compound by passage of thecompound through the mucosal tissue and into the blood stream.

[0038] “Penetration enhancement” or “permeation enhancement” refers toan increase in the permeability of the skin or mucosal tissue to aselected pharmacologically active compound such that the rate at whichthe compound permeates through the skin or mucosal tissue is increased.

[0039] “Vaginal delivery” refers to the direct administration of apharmaceutical composition to the vagina of the patient. Generally,“vaginal delivery” of a pharmaceutical composition involvesadministration to the distal several centimeters of the vagina.

[0040] “Vulvar delivery” or “vulvar administration” to refer toapplication of a pharmaceutical composition to the vulvar area of apatient. The term is intended to encompass application to the clitorisas well as the surrounding vulvar area. The terms “vulvar delivery” and“clitoral delivery” are used interchangeably herein and are bothintended to refer to administration to the vulvar area of the patient.

[0041] “Carriers” or “vehicles” refers to carrier materials suitable forcompound administration and include any such material known in the artsuch as, for example, any liquid, gel, solvent, liquid diluent,solubilizer, or the like, which is non-toxic and which does not interactwith any components of the composition in a deleterious manner.

[0042] “Sustained release” refers to the release of a therapeuticallyactive compound and/or composition such that the blood levels of thetherapeutically active compound are maintained within a desirabletherapeutic range over an extended period of time. The sustained releaseformulation can be prepared using any conventional method known to oneskilled in the art to obtain the desired release characteristics.

[0043] “Nitric oxide donor” or “NO donor” refers to compounds thatdonate, release and/or directly or indirectly transfer a nitrogenmonoxide species, and/or stimulate the endogenous production of nitricoxide or endothelium-derived relaxing factor (EDRF) in vivo and/orelevate endogenous levels of nitric oxide or EDRF in vivo. “NO donor”also includes compounds that are substrates for nitric oxide synthase.

[0044] “Nitric oxide adduct” or “NO adduct” refers to compounds andfunctional groups which, under physiological conditions, can donate,release and/or directly or indirectly transfer any of the three redoxforms of nitrogen monoxide (NO⁺, NO⁻, NO), such that the biologicalactivity of the nitrogen monoxide species is expressed at the intendedsite of action.

[0045] “Nitric oxide releasing” or “nitric oxide donating” refers tomethods of donating, releasing and/or directly or indirectlytransferring any of the three redox forms of nitrogen monoxide (NO⁺,NO⁻, NO), such that the biological activity of the nitrogen monoxidespecies is expressed at the intended site of action.

[0046] “Alkyl” refers to a lower alkyl group, a haloalkyl group, analkenyl group, an alkynyl group, a bridged cycloalkyl group, acycloalkyl group or a heterocyclic ring, as defined herein.

[0047] “Lower alkyl” refers to branched or straight chain acyclic alkylgroup comprising one to about ten carbon atoms (preferably one to abouteight carbon atoms, more preferably one to about six carbon atoms).Exemplary lower alkyl groups include methyl, ethyl, n-propyl, isopropyl,n-butyl, isobutyl, sec-butyl, t-butyl, pentyl, neopentyl, iso-amyl,hexyl, octyl, and the like.

[0048] “Haloalkyl” refers to a lower alkyl group, an alkenyl group, analkynyl group, a bridged cycloalkyl group, a cycloalkyl group or aheterocyclic ring, as defined herein, to which is appended one or morehalogens, as defined herein. Exemplary haloalkyl groups includetrifluoromethyl, chloromethyl, 2-bromobutyl, 1-bromo-2-chloro-pentyl,and the like.

[0049] “Alkenyl” refers to a branched or straight chain C₂-C₁₀hydrocarbon (preferably a C₂-C₈ hydrocarbon, more preferably a C₂-C₆hydrocarbon) which can comprise one or more carbon-carbon double bonds.Exemplary alkenyl groups include propylenyl, buten-1-yl, isobutenyl,penten-1-yl, 2,2-methylbuten-1-yl, 3-methylbuten-1-yl, hexan-1-yl,hepten-1-yl, octen-1-yl, and the like.

[0050] “Alkynyl” refers to an unsaturated acyclic C₂-C₁₀ hydrocarbon(preferably a C₂-C₈ hydrocarbon, more preferably a C₂-C₆ hydrocarbon)which can comprise one or more carbon-carbon triple bonds. Exemplaryalkynyl groups include ethynyl, propynyl, butyn-1-yl, butyn-2-yl,pentyl-1-yl, pentyl-2-yl, 3-methylbutyn-1-yl, hexyl-1-yl, hexyl-2-yl,hexyl-3-yl, 3,3-dimethyl-butyn-1-yl, and the like.

[0051] “Bridged cycloalkyl” refers to two or more cycloalkyl groups,heterocyclic groups, or a combination thereof fused via adjacent ornon-adjacent atoms. Bridged cycloalkyl groups can be unsubstituted orsubstituted with one, two or three substituents independently selectedfrom alkyl, alkoxy, amino, alkylamino, dialkylamino, hydroxy, halo,carboxyl, alkylcarboxylic acid, aryl, amidyl, ester, alkylcarboxylicester, carboxamido, alkylcarboxamido, oxo and nitro. Exemplary bridgedcycloalkyl groups include adamantyl, decahydronapthyl, quinuclidyl,2,6-dioxabicyclo(3.3.0)octane, 7-oxabycyclo(2.2.1)heptyl,8-azabicyclo(3,2, 1)oct-2-enyl and the like.

[0052] “Cycloalkyl” refers to a saturated or unsaturated cyclichydrocarbon comprising from about 3 to about 10 carbon atoms. Cycloalkylgroups can be unsubstituted or substituted with one, two or threesubstituents independently selected from alkyl, alkoxy, amino,alkylamino, dialkylamino, arylamino, diarylamino, alkylarylamino, aryl,amidyl, ester, hydroxy, halo, carboxyl, alkylcarboxylic acid,alkylcarboxylic ester, carboxamido, alkylcarboxamido, oxo and nitro.Exemplary cycloalkyl groups include cyclopropyl, cyclobutyl,cyclopentyl, cyclohexyl, cyclohexenyl, cyclohepta, 1,3-dienyl, and thelike.

[0053] “Heterocyclic ring or group” refers to a saturated, unsaturated,cyclic or aromatic or polycyclic hydrocarbon group having about 2 toabout 10 carbon atoms (preferably about 4 to about 6 carbon atoms) where1 to about 4 carbon atoms are replaced by one or more nitrogen, oxygenand/or sulfur atoms. Sulfur may be in the thio, sulfinyl or sulfonyloxidation state. The heterocyclic ring or group can be fused to anaromatic hydrocarbon group. Heterocyclic groups can be unsubstituted orsubstituted with one, two or three substituents independently selectedfrom alkyl, alkoxy, amino, alkylamino, dialkylamino, arylamino,diarylamino, alkylarylamino, hydroxy, oxo, thial, halo, carboxyl,carboxylic ester, alkylcarboxylic acid, alkylcarboxylic ester, aryl,arylcarboxylic acid, arylcarboxylic ester, amidyl, ester, carboxamido,alkylcarboxamido, arylcarboxamido, sulfonic acid, sulfonic ester,sulfonamido and nitro. Exemplary heterocyclic groups include pyrrolyl,3-pyrrolinyl,4,5,6-trihydro-2H-pyranyl, pyridinyl, 1,4-dihydropyridinyl,pyrazolyl, triazolyl, pyrimidinyl, pyridazinyl, oxazolyl, thiazolyl,imidazolyl, indolyl, thiophenyl, furanyl, tetrhydrofuranyl, tetrazolyl,2-pyrrolinyl, 3-pyrrolinyl, pyrrolindinyl, oxazolindinyl 1,3-dioxolanyl,2,6-dioxabicyclo(3,3,0)octanyl, 2-imidazonlinyl, imidazolindinyl,2-pyrazolinyl, pyrazolidinyl, isoxazolyl, isothiazolyl,1,2,3-oxadiazolyl, 1,2,3-triazolyl, 1,3,4-thiadiazolyl, 2H-pyranyl,4H-pyranyl, piperidinyl, 1,4-dioxanyl, morpholinyl, 1,4-dithianyl,thiomorpholinyl, pyrazinyl, piperazinyl, 1,3,5-triazinyl,1,3,5-trithianyl, benzo(b)thiophenyl, benzimidazolyl, quinolinyl, andthe like.

[0054] “Heterocyclic compounds” refer to mono- and polycyclic compoundscomprising at least one aryl or heterocyclic ring.

[0055] “Aryl” refers to a monocyclic, bicyclic, carbocyclic orheterocyclic ring system comprising one or two aromatic rings. Exemplaryaryl groups include phenyl, pyridyl, napthyl, quinoyl,tetrahydronaphthyl, furanyl, indanyl, indenyl, indoyl, and the like.Aryl groups (including bicylic aryl groups) can be unsubstituted orsubstituted with one, two or three substituents independently selectedfrom alkyl, alkoxy, amino, alkylamino, dialkylamino, arylamino,diarylamino, alkylarylamino, hydroxy, carboxyl, carboxylic ester,alkylcarboxylic acid, alkylcarboxylic ester, aryl, arylcarboxylic acid,arylcarboxylic ester, alkylcarbonyl, arylcarbonyl, amidyl, ester,carboxamido, alkylcarboxamido, carbomyl, sulfonic acid, sulfonic ester,sulfonamido and nitro. Exemplary substituted aryl groups includetetrafluorophenyl, pentafluorophenyl, sulfonamide, alkylsulfonyl,arylsulfonyl, and the like.

[0056] “Alkylaryl” refers to an alkyl group, as defined herein, to whichis appended an aryl group, as defined herein. Exemplary alkylaryl groupsinclude benzyl, phenylethyl, hydroxybenzyl, fluorobenzyl,fluorophenylethyl, and the like.

[0057] “Arylalkyl” refers to an aryl radical, as defined herein,attached to an alkyl radical, as defined herein.

[0058] “Cycloalkylalkyl” refers to a cycloalkyl radical, as definedherein, attached to an alkyl radical, as defined herein.

[0059] “Heterocyclicalkyl” refers to a heterocyclic ring radical, asdefined herein, attached to an alkyl radical, as defined herein.

[0060] “Cycloalkenyl” refers to an unsaturated cyclic hydrocarbon havingabout 3 to about 10 carbon atoms (preferably about 3 to about 8 carbonatoms, more preferably about 3 to about 6 carbon atoms) comprising oneor more carbon-carbon double bonds.

[0061] “Arylheterocyclic ring” refers to a bi- or tricyclic ringcomprised of an aryl ring, as defined herein, appended via two adjacentcarbon atoms of the aryl ring to a heterocyclic ring, as defined herein.Exemplary arylheterocyclic rings include dihydroindole,1,2,3,4-tetra-hydroquinoline, and the like.

[0062] “Alkoxy” refers to R₅₀O—, wherein R₅₀ is an alkyl group, asdefined herein. Exemplary alkoxy groups include methoxy, ethoxy,t-butoxy, cyclopentyloxy, and the like.

[0063] “Arylalkoxy or alkoxyaryl” refers to an alkoxy group, as definedherein, to which is appended an aryl group, as defined herein. Exemplaryarylalkoxy groups include benzyloxy, phenylethoxy, chlorophenylethoxy,and the like.

[0064] “Aryloxy” refers to R₅₅O—, wherein R₅₅ is an aryl group, asdefined herein. Exemplary aryloxy groups include napthyloxy,quinolyloxy, isoquinolizinyloxy, and the like.

[0065] “Alkoxyalkyl” refers to an alkoxy group, as defined herein,appended to an alkyl group, as defined herein. Exemplary alkoxyalkylgroups include methoxymethyl, methoxyethyl, isopropoxymethyl, and thelike.

[0066] “Alkoxyhaloalkyl” refers to an alkoxy group, as defined herein,appended to a haloalkyl group, as defined herein. Exemplaryalkoxyhaloalkyl groups include 4-methoxy-2-chlorobutyl and the like.

[0067] “Cycloalkoxy” refers to R₅₄O—, wherein R₅₄ is a cycloalkyl groupor a bridged cycloalkyl group, as defined herein. Exemplary cycloalkoxygroups include cyclopropyloxy, cyclopentyloxy, cyclohexyloxy, and thelike.

[0068] “Haloalkoxy” refers to a haloalkyl group, as defined herein, towhich is appended an alkoxy group, as defined herein. Exemplaryhaloalkyl groups include 1,1,1-trichloroethoxy, 2-bromobutoxy, and thelike.

[0069] “Hydroxy” refers to —OH.

[0070] “Oxo” refers to ═O.

[0071] “Oxy” refers to —O.R₇₇ ⁺ wherein R₇₇ is an organic or inorganiccation.

[0072] “Organic cation” refers to a positively charged organic ion.Exemplary organic cations include alkyl substituted ammonium cations,and the like.

[0073] “Inorganic cation” refers to a positively charged metal ion.Exemplary inorganic cations include Group I metal cations such as forexample, sodium, potassium, and the like.

[0074] “Hydroxyalkyl” refers to a hydroxy group, as defined herein,appended to an alkyl group, as defined herein.

[0075] “Amino” refers to —NH₂.

[0076] “Nitrate” refers to —O—NO₂.

[0077] “Nitrite” refers to —O—NO.

[0078] “Thionitrate” refers to —S—NO₂.

[0079] “Thionitrite” and “nitrosothiol” refer to —S—NO.

[0080] “Nitro” refers to the group —NO₂ and “nitrosated” refers tocompounds that have been substituted therewith.

[0081] “Nitroso” refers to the group —NO and “nitrosylated” refers tocompounds that have been substituted therewith.

[0082] “Nitrile” and “cyano” refer to —CN.

[0083] “Halogen” or “halo” refers to iodine (I), bromine (Br), chlorine(Cl), and/or fluorine (F).

[0084] “Alkylamino” refers to R₅₀NH—, wherein R₅₀ is an alkyl group, asdefined herein. Exemplary alkylamino groups include methylamino,ethylamino, butylamino, cyclohexylamino, and the like.

[0085] “Arylamino” refers to R₅₅NH—, wherein R₅₅ is an aryl group, asdefined herein.

[0086] “Dialkylamino” refers to R₅₀R₅₂N—, wherein R₅₀ and R₅₂ are eachindependently an alkyl group, as defined herein. Exemplary dialkylaminogroups include dimethylamino, diethylamino, methyl propargylamino, andthe like.

[0087] “Diarylamino” refers to R₅₅R₆₀N—, wherein R₅₅ and R₆₀ are eachindependently an aryl group, as defined herein.

[0088] “Alkylarylamino” refers to R₅₀R₅₅N—, wherein R₅₀ is an alkylgroup, as defined herein, and R₅₅ is an aryl group, as defined herein.

[0089] “Aminoalkyl” refers to an amino group, an alkylamino group, adialkylamino group, an arylamino group, a diarylamino group, analkylarylamino group or a heterocyclic ring, as defined herein, to whichis appended an alkyl group, as defined herein.

[0090] “Aminoaryl ” refers to an amino group, an alkylamino group, adialkylamino group, an arylamino group, a diarylamino group, analkylarylamino group or a heterocyclic ring, as defined herein, to whichis appended an aryl group, as defined herein.

[0091] “Thio” refers to —S—.

[0092] “Sulfinyl” refers to —S(O)—.

[0093] “Methanthial” refers to —C(S)—.

[0094] “Thial” refers to ═S.

[0095] “Sulfonyl” refers to —S(O)₂ ⁻.

[0096] “Sulfonic acid” refers to —S(O)₂OR₇₆, wherein R₇₆ is a hydrogen,an organic cation or an inorganic cation.

[0097] “Alkylsulfonic acid” refers to a sulfonic acid group, as definedherein, appended to an alkyl group, as defined herein.

[0098] “Arylsulfonic acid” refers to an sulfonic acid group, as definedherein, appended to an aryl group, as defined herein

[0099] “Sulfonic ester” refers to —S(O)₂OR₅₈, wherein R₅₈ is an alkylgroup, an aryl group, an alkylaryl group or an aryl heterocyclic ring,as defined herein.

[0100] “Sulfonamido” refers to —S(O)₂—N(R₅₁)(R₅₇), wherein R₅, and R₅₇are each independently a hydrogen atom, an alkyl group, an aryl group,an alkylaryl group, or an arylheterocyclic ring, as defined herein, orR₅₁ and R₅₇ taken together are a heterocyclic ring, a cycloalkyl groupor a bridged cycloalkyl group, as defined herein.

[0101] “Alkylsulfonamido” refers to a sulfonamido group, as definedherein, appended to an alkyl group, as defined herein.

[0102] “Arylsulfonamido” refers to a sulfonamido group, as definedherein, appended to an aryl group, as defined herein.

[0103] “Alkylthio” refers to R₅₀S—, wherein R₅₀ is an alkyl group, asdefined herein.

[0104] “Arylthio” refers to R₅₅S—, wherein R₅₅ is an aryl group, asdefined herein.

[0105] “Cycloalkylthio” refers to R₅₄S—, wherein R₅₄ is a cycloalkylgroup or a bridged cycloalkyl group, as defined herein. Exemplarycycloalkylthio groups include cyclopropylthio, cyclopentylthio,cyclohexylthio, and the like.

[0106] “Alkylsulfinyl” refers to R₅₀—S(O)—, wherein R₅₀ is an alkylgroup, as defined herein.

[0107] “Alkylsulfonyl” refers to R₅₀—S(O)₂—, wherein R₅₀ is an alkylgroup, as defined herein.

[0108] “Arylsulfinyl” refers to R₅₅—S(O)—, wherein R₅₅ is an aryl group,as defined herein.

[0109] “Arylsulfonyl” refers to R₅₅—S(O)₂—, wherein R₅₅ is an arylgroup, as defined herein.

[0110] “Amidyl” refers to R₅₁C(O)N(R₅₇)— wherein R₅₁ and R₅₇ are eachindependently a hydrogen atom, an alkyl group, an aryl group, analkylaryl group, or an arylheterocyclic ring, as defined herein.

[0111] “Ester” refers to R₅₁C(O)O— wherein R₅₁ is a hydrogen atom, analkyl group, an aryl group, an alkylaryl group, or an arylheterocyclicring, as defined herein.

[0112] “Carbamoyl” refers to —O—C(O)N(R₅₁)(R₅₇), wherein R₅₁ and R₅₇ areeach independently a hydrogen atom, an alkyl group, an aryl group, analkylaryl group or an arylheterocyclic ring, as defined herein, or R₅₁and R₅₇ taken together are a heterocyclic ring, a cycloalkyl group or abridged cycloalkyl group, as defined herein.

[0113] “Carbamate” refers to R₅₁O—C(O)N—(R₅₇), wherein R₅₁ and R₅₇ areeach independently a hydrogen atom, an alkyl group, an aryl group, analkylaryl group or an arylheterocyclic ring, as defined herein, or R₅₁and R₅₇ taken together are a heterocyclic ring, a cycloalkyl group or abridged cycloalkyl group, as defined herein.

[0114] “Carboxyl” refers to —C(O)OR₇₆, wherein R₇₆ is a hydrogen, anorganic cation or an inorganic cation, as defined herein.

[0115] “Carbonyl” refers to —C(O)—.

[0116] “Alkylcarbonyl” or “alkanoyl” refers to R₅₀—C(O)—, wherein R₅₀ isan alkyl group, as defined herein.

[0117] “Arylcarbonyl” or “aroyl” refers to R₅₅—C(O)—, wherein R₅₅ is anaryl group, as defined herein.

[0118] “Carboxylic ester” refers to —C(O)OR₅₈, wherein R₅₈ is an alkylgroup, an aryl group, an alkylaryl group or an aryl heterocyclic ring,as defined herein.

[0119] “Alkylcarboxylic acid” and “alkylcarboxyl” refer to an alkylgroup, as defined herein, appended to a carboxyl group, as definedherein.

[0120] “Alkylcarboxylic ester” refers to an alkyl group, as definedherein, appended to a carboxylic ester group, as defined herein.

[0121] “Arylcarboxylic acid” refers to an aryl group, as defined herein,appended to a carboxyl group, as defined herein.

[0122] “Arylcarboxylic ester” and “arylcarboxyl” refer to an aryl group,as defined herein, appended to a carboxylic ester group, as definedherein.

[0123] “Carboxamido” refers to —C(O)N(R₅₁)(R₅₇), wherein R₅₁ and R₅₇ areeach independently a hydrogen atom, an alkyl group, an aryl group, analkylaryl group or an arylheterocyclic ring, as defined herein, or R₅₁and R₅₇ taken together with the nitrogen to which they are attached forma heterocyclic ring, a cycloalkyl group or a bridged cycloalkyl group,as defined herein.

[0124] “Alkylcarboxamido” refers to an alkyl group, as defined herein,appended to a carboxamido group, as defined herein.

[0125] “Arylcarboxamido” refers to an aryl group, as defined herein,appended to a carboxamido group, as defined herein.

[0126] “Urea” refers to —N(R₅₉)—C(O)N(R₅₁)(R₅₇) wherein R₅₁, R₅₇, andR₅₉ are each independently a hydrogen atom, an alkyl group, an arylgroup, an alkylaryl group, or an arylheterocyclic ring, as definedherein, or R₅₁ and R₅₇ taken together with the nitrogen to which theyare attached form a heterocyclic ring, as defined herein.

[0127] “Phosphoryl” refers to —P(R₇₀)(R₇₁)(R₇₂), wherein R₇₀ is a lonepair of electrons, sulfur or oxygen, and R₇₁ and R₇₂ are eachindependently a covalent bond, a hydrogen, a lower alkyl, an alkoxy, analkylamino, a hydroxy or an aryl, as defined herein.

[0128] “Silyl” refers to —Si(R₇₃)(R₇₄)(R₇₅), wherein R₇₃, R₇₄ and R₇₅are each independently a covalent bond, a lower alkyl, an alkoxy, anaryl or an arylalkoxy, as defined herein.

[0129] The term “sexual dysfunction” generally includes any sexualdysfunction in a patient, including an animal, preferably a mammal, morepreferably a human. The patient can be male or female. Sexualdysfunctions can include, for example, sexual desire disorders, sexualarousal disorders, orgasmic disorders and sexual pain disorders. Femalesexual dysfunction refers to any female sexual dysfunction including,for example, sexual desire disorders, sexual arousal dysfunctions,orgasmic dysfunctions, sexual pain disorders, dyspareunia, andvaginismus. The female can be pre-menopausal or menopausal. Sexualdysfunction can be caused, for example, by pregnancy, menopause, cancer,pelvic surgery, chronic medical illness or medications. Male sexualdysfunction refers to any male sexual dysfunctions including, forexample, male erectile dysfunction and impotence.

[0130] The present invention is directed to methods for measuringvasodilation and changes in blood flow in patients following theadministration of a nitric oxide donor and/or vasoactive agent usinginfrared thermography. The monitoring and diagnosing of diseases anddisorders related to vasodilation and changes in blood flow, such as,for example, sexual dysfunctions in patients, including males andfemales, by administering the nitric oxide donors and/or vasoactiveagents are also described herein.

[0131] A principal aspect of the present invention relates to measuringthe temperature of an area of interest using infrared thermography priorto and/or during and/or following administration of at least one nitricoxide donor and/or at least one vasoactive agent. Any infraredthermographic imaging system known to one skilled in the art can be usedin the present invention. For example, THERMACAM® X90 infrared focalplane array (FPA) series, THERMACAM SC 1000 series and THERMACAM SC 3000are available from Inframetrics, Inc., North Billerica, Mass.; AGEMAseries are commercially available from FLIR Systems, Inc., Portland,Oreg.; WO 99/10731 discloses the use of microwave enhanced infraredthermography for the detection, location and identification of objects;and WO 99/60630 discloses the use of infrared thermographic imagingsystems to monitor the physiological and molecular thermogenic events inanimals, plants, tissues and isolated cells; the disclosures of each ofwhich is incorporated by reference herein in its entirety.

[0132] One embodiment of the present invention comprises methods formonitoring and/or measuring the vasodilation and changes in blood flowin vasculature using infrared thermography resulting from theadministration of at least one compound that donates, transfers orreleases nitric oxide, elevates endogenous levels of endothelium-derivedrelaxing factor, or EDRF in vivo, stimulates endogenous synthesis ofnitric oxide or is a substrate for nitric oxide synthase, and/orotherwise directly or indirectly deliver or transfer nitric oxide to asite of its activity, such as on a cell membrane, in vivo (i.e., nitricoxide donor). For example, the method for monitoring and measuringvasodilation and changes in blood flow may comprise (i) measuring thebaseline temperature of an area of interest using infrared thermography,(ii) administering to a patient at least one nitric oxide donor, (iii)measuring the temperature of the area of interest during and/or afteradministering to a patient the at least one nitric oxide donor usinginfrared thermography, and (iv) comparing the measurements obtained insteps (i) and (iii), wherein a nitric oxide donor that results in anincrease in the temperature between step (i) and step (iii) is avasodilator and a nitric oxide donor that results in a decrease in thetemperature between step (i) and step (iii) is a vasoconstrictor. Inanother embodiment, a plurality of nitric oxide donors can be tested inthe methods of the present invention, and the results of each test canbe compared to determine which nitric oxide donor is the most effective,i.e., which nitric oxide donor produces the greatest increase intemperature between step (i) and step (iii). The baseline temperature instep (i) may be a previously obtained measurement; or alternatively ameasurement taken after the administration of the compound when a stablebaseline temperature measurement is obtained. The nitric oxide donor canoptionally be administered with at least one vasoactive agent.Contemplated nitric oxide donors and vasoactive agents include all thoseknown in the art and those described herein. The nitric oxide donorsand/or vasoactive agents can be administered separately or in the formof a composition. The compounds and compositions of the presentinvention can also be administered in combination with other medicationsused for monitoring diseases or disorders.

[0133] As used herein, the term “nitric oxide” encompasses unchargednitric oxide (NO.) and charged nitrogen monoxide species, preferablycharged nitrogen monoxide species, such as nitrosonium ion (NO⁺) andnitroxyl ion (NO⁻). NO. is a highly reactive short-lived species that ispotentially toxic to cells. This is critical because the pharmacologicalefficacy of NO depends upon the form in which it is delivered. Incontrast to the nitric oxide radical (NO.), nitrosonium (NO⁺) does notreact with O₂ or O₂ ⁻ species, and functionalities capable oftransferring and/or releasing NO⁺ and NO− are also resistant todecomposition in the presence of many redox metals.

[0134] The term “nitric oxide” encompasses uncharged nitric oxide (NO.)and charged nitrogen monoxide species, preferably charged nitrogenmonoxide species, such as nitrosonium ion (NO⁺) and nitroxyl ion (NO−).The reactive form of nitric oxide can be provided by gaseous nitricoxide. The nitrogen monoxide releasing, delivering or transferringcompounds include any and all such compounds which provide nitrogenmonoxide to its intended site of action in a form active for itsintended purpose. The term “NO adducts” encompasses any nitrogenmonoxide releasing, delivering or transferring compounds, including, forexample, S-nitrosothiols, nitrites, nitrates, S-nitrothiols,sydnonimines, 2-hydroxy-2-nitrosohydrazines (NONOates),(E)-alkyl-2-((E)-hydroxyimino)-5-nitro-3-hexene amines or amides,nitrosoamines, furoxans as well as substrates for the endogenous enzymeswhich synthesize nitric oxide. NONOates include, but are not limited to,(Z)-1-{N-methyl-N-(6-(N-methyl-ammoniohexyl)amino)}diazen-1-ium-1,2-diolate (“MAHMA/NO”),(Z)-1-(N-(3-ammoniopropyl)-N-(n-propyl)amino)diazen-1-ium-1,2-diolate(“PAPA/NO”),(Z)-1-{N-(3-aminopropyl)-N-(4-(3-aminopropylammonio)butyl)-amino}diazen-1-ium-1,2-diolate(spermine NONOate or “SPER/NO”) and sodium(Z)-1-(N,N-diethylamino)diazenium-1,2-diolate (diethylamine NONOate or“DEA/NO”) and derivatives thereof. The “NO adducts” can bemono-nitrosylated, poly-nitrosylated, mono-nitrosated and/orpoly-nitrosated or a combination thereof at a variety of naturallysusceptible or artificially provided binding sites for biologicallyactive forms of nitrogen monoxide.

[0135] One group of NO adducts is the S-nitrosothiols, which arecompounds that include at least one —S—NO group. These compounds includeS-nitroso-polypeptides (the term “polypeptide” includes proteins andpolyamino acids that do not possess an ascertained biological function,and derivatives thereof); S-nitrosylated amino acids (including naturaland synthetic amino acids and their stereoisomers and racemic mixturesand derivatives thereof); S-nitrosylated sugars; S-nitrosylated,modified and unmodified, oligonucleotides (preferably of at least 5, andmore preferably 5-200 nucleotides); straight or branched, saturated orunsaturated, aliphatic or aromatic, substituted or unsubstitutedS-nitrosylated hydrocarbons; and S-nitroso heterocyclic compounds.S-nitrosothiols and methods for preparing them are described in U.S.Pat. Nos. 5,380,758 and 5,703,073; WO 97/27749; WO 98/19672; and Oae etal, Org. Prep. Proc. Int., 15(3): 165-198 (1983), the disclosures ofeach of which are incorporated by reference herein in their entirety.

[0136] Another embodiment of the present invention is S-nitroso aminoacids where the nitroso group is linked to a sulfur group of asulfur-containing amino acid or derivative thereof. Such compoundsinclude, for example, S-nitroso-N-acetylcysteine, S-nitroso-captopril,S-nitroso-N-acetylpenicillamine, S-nitroso-homocysteine,S-nitroso-cysteine, S-nitroso-glutathione andS-nitroso-cysteinyl-glycine. In a preferred embodiment, the S-nitrosoamino acid is S-nitroso-glutathione.

[0137] Suitable S-nitrosylated proteins include thiol-containingproteins (where the NO group is attached to one or more sulfur groups onan amino acid or amino acid derivative thereof) from various functionalclasses including enzymes, such as tissue-type plasminogen activator(TPA) and cathepsin B; transport proteins, such as lipoproteins; hemeproteins, such as hemoglobin and serum albumin; and biologicallyprotective proteins, such as immunoglobulins, antibodies and cytokines.Such nitrosylated proteins are described in WO 93/09806, the disclosureof which is incorporated by reference herein in its entirety. Examplesinclude polynitrosylated albumin where one or more thiol or othernucleophilic centers in the protein are modified.

[0138] Other examples of suitable S-nitrosothiols include:

[0139] (i) HS(C(R_(e))(R_(f)))_(m)SNO;

[0140] (ii) ONS(C(R_(e))(R_(f)))_(m)R_(e); and

[0141] (iii) H₂N—CH(CO₂H)—(CH₂)_(m)—C(O)NH—CH(CH₂SNO)—C(O)NH—CH₂—CO₂H;

[0142] wherein m is an integer from 2 to 20; R_(e) and R_(f) are eachindependently a hydrogen, an alkyl, a cycloalkoxy, a halogen, a hydroxy,an hydroxyalkyl, an alkoxyalkyl, an arylheterocyclic ring, an alkylaryl,a cycloalkylalkyl, a heterocyclicalkyl, an alkoxy, a haloalkoxy, anamino, an alkylamino, a dialkylamino, an arylamino, a diarylamino, analkylarylamino, an alkoxyhaloalkyl, a haloalkoxy, a sulfonic acid, asulfonic ester, an alkylsulfonic acid, an arylsulfonic acid, anarylalkoxy, an alkylthio, an arylthio, a cycloalkylthio, a cycloalkenyl,a cyano, an aminoalkyl, an aminoaryl, an aryl, an arylalkyl, analkylaryl, a carboxamido, a alkylcarboxamido, an arylcarboxamido, anamidyl, a carboxyl, a carbamoyl, a carbamate, an alkylcarboxylic acid,an arylcarboxylic acid, an alkylcarbonyl, an arylcarbonyl, an ester, acarboxylic ester, an alkylcarboxylic ester, an arylcarboxylic ester, ahaloalkoxy, a sulfonamido, an alkylsulfonamido, an arylsulfonamido, asulfonic ester, a urea, a phosphoryl, a nitro, -T-Q, or—(C(R_(e))(R_(f)))_(k)-T-Q, or R_(e) and R_(f) taken together with thecarbons to which they are attached form a carbonyl, a methanthial, aheterocyclic ring, a cycloalkyl group or a bridged cycloalkyl group; Qis —NO or —NO₂; and T is independently a covalent bond, a carbonyl, anoxygen, —S(O)_(o)— or —N(R_(a))R_(i)—, wherein o is an integer from 0 to2, R_(a) is a lone pair of electrons, a hydrogen or an alkyl group;R_(i) is a hydrogen, an alkyl, an aryl, an alkylcarboxylic acid, an arylcarboxylic acid, an alkylcarboxylic ester, an arylcarboxylic ester, analkylcarboxamido, an arylcarboxamido, an alkylaryl, an alkylsulfinyl, analkylsulfonyl, an arylsulfinyl, an arylsulfonyl, a sulfonamido, acarboxamido, a carboxylic ester, an amino alkyl, an amino aryl,—CH₂—C(T-Q)(R_(e))(R_(f)), or —(N₂O₂—)⁻.M⁺, wherein M⁺ is an organic orinorganic cation; with the proviso that when R₁ is—CH₂—C(T-Q)(R_(e))(R_(f)) or

[0143] —(N₂O₂—).M⁺; then “-T-Q” can be a hydrogen, an alkyl group, analkoxyalkyl group, an aminoalkyl group, a hydroxy group or an arylgroup.

[0144] In cases where R_(e) and R_(f) are a heterocyclic ring or R_(e)and R_(f) when taken together with the carbon atoms to which they areattached are a heterocyclic ring, then R_(i) can be a substituent on anydisubstituted nitrogen contained within the radical wherein R_(i) is asdefined herein.

[0145] Nitrosothiols can be prepared by various methods of synthesis. Ingeneral, the thiol precursor is prepared first, then converted to theS-nitrosothiol derivative by nitrosation of the thiol group with NaNO₂under acidic conditions (pH is about 2.5) which yields the S-nitrosoderivative. Acids which can be used for this purpose include aqueoussulfuric, acetic and hydrochloric acids. The thiol precursor can also benitrosylated by reaction with an organic nitrite such as tert-butylnitrite, or a nitrosonium salt such as nitrosonium tetraflurorborate inan inert solvent.

[0146] Another group of NO adducts for use in the present invention,where the NO adduct is a compound that donates, transfers or releasesnitric oxide, include compounds comprising at least one ON—O—, ON—N— orON—C— group. The compounds that include at least one ON—O—, ON—N— orON—C— group are preferably ON—O—, ON—N— or ON—C-polypeptides (the term“polypeptide” includes proteins and polyamino acids that do not possessan ascertained biological function, and derivatives thereof); ON—O,ON—N— or ON—C-amino acids (including natural and synthetic amino acidsand their stereoisomers and racemic mixtures); ON—O—, ON—N— orON—C-sugars; ON—O—, ON—N— or ON—C— modified or unmodifiedoligonucleotides (comprising at least 5 nucleotides, preferably 5-200nucleotides); ON—O—, ON—N— or ON—C— straight or branched, saturated orunsaturated, aliphatic or aromatic, substituted or unsubstitutedhydrocarbons; and ON—O—, ON—N— or ON—C-heterocyclic compounds.

[0147] Another group of NO adducts for use in the present inventioninclude nitrates that donate, transfer or release nitric oxide, such ascompounds comprising at least one O₂N—O—, O₂N—N—, O₂N—S— or O₂N—C—group. Preferred among these compounds are O₂N—O—, O₂N—N—, O₂N—S— orO₂N—C— polypeptides (the term “polypeptide” includes proteins and alsopolyamino acids that do not possess an ascertained biological function,and derivatives thereof); O₂N—O—, O₂N—N—, O₂N—S— or O₂N—C— amino acids(including natural and synthetic amino acids and their stereoisomers andracemic mixtures); O₂N—O—, O₂N—N—, O₂N—S— or O₂N—C-sugars; O₂N—O—,O₂N—N—, O₂N—S— or O₂N—C— modified and unmodified oligonucleotides(comprising at least 5 nucleotides, preferably 5-200 nucleotides);O₂N—O—, O₂N—N—, O₂N—S— or O₂N—C— straight or branched, saturated orunsaturated, aliphatic or aromatic, substituted or unsubstitutedhydrocarbons; and O₂N—O—, O₂N—N—, O₂N—S— or O₂N—C— heterocycliccompounds. Preferred examples of compounds comprising at least oneO₂N—O—, O₂N—N—, O₂N—S— or O₂N—C— group include isosorbide dinitrate,isosorbide mononitrate, clonitrate, erythrityltetranitrate, mannitolhexanitrate, nitroglycerin, pentaerythritoltetranitrate, pentrinitroland propatylnitrate.

[0148] Another group of NO adducts are N-oxo-N-nitrosoamines thatdonate, transfer or release nitric oxide and are represented by theformula: R¹R²—N(O-M⁺)—NO, where R¹ and R² are each independently apolypeptide, an amino acid, a sugar, a modified or unmodifiedoligonucleotide, a straight or branched, saturated or unsaturated,aliphatic or aromatic, substituted or unsubstituted hydrocarbon, or aheterocyclic group, and M⁺ is as defined herein.

[0149] Another group of NO adducts are thionitrates that donate,transfer or release nitric oxide and are represented by the formula:R¹—(S)—NO₂, where R¹ is a polypeptide, an amino acid, a sugar, amodified or unmodified oligonucleotide, a straight or branched,saturated or unsaturated, aliphatic or aromatic, substituted orunsubstituted hydrocarbon, or a heterocyclic group. Preferred are thosecompounds where R¹ is a polypeptide or hydrocarbon with a pair or pairsof thiols that are sufficiently structurally proximate, i.e., vicinal,that the pair of thiols will be reduced to a disulfide. Compounds whichform disulfide species release nitroxyl ion (NO−) and uncharged nitricoxide (NO.).

[0150] The present invention is also directed to compounds thatstimulate endogenous NO or elevate levels of endogenousendothelium-derived relaxing factor (EDRF) in vivo or are substrates forthe enzyme, nitric oxide synthase. Such compounds include, for example,L-arginine, L-homoarginine, and N-hydroxy-L-arginine, including theirnitrosated and nitrosylated analogs (e.g., nitrosated L-arginine,nitrosylated L-arginine, nitrosated N-hydroxy-L-arginine, nitrosylatedN-hydroxy-L-arginine, nitrosated L-homoarginine and nitrosylatedL-homoarginine), precursors of L-arginine and/or physiologicallyacceptable salts thereof, including, for example, citrulline, ornithine,glutamine, lysine, polypeptides comprising at least one of these aminoacids, inhibitors of the enzyme arginase (e.g., N-hydroxy-L-arginine and2(S)-amino-6-boronohexanoic acid) and the substrates for nitric oxidesynthase, cytokines, adenosin, bradykinin, calreticulin, bisacodyl, andphenolphthalein. EDRF is a vascular relaxing factor secreted by theendothelium, and has been identified as nitric oxide (NO) or a closelyrelated derivative thereof (Palmer et al, Nature, 327:524-526 (1987);Ignarro et al, Proc. Natl. Acad. Sci. USA, 84:9265-9269 (1987)).

[0151] Another embodiment of the present invention comprises methods formonitoring and/or measuring the vasodilation and changes in blood flowin vasculature using infrared thermography resulting from theadministration of at least one vasoactive agent. For example, the methodfor monitoring and measuring vasodilation and changes in blood flow maycomprise (i) measuring the baseline temperature of an area of interestusing infrared thermography, (ii) administering to a patient at leastone vasoactive agent, (iii) measuring the temperature of the area ofinterest during and/or after administering the at least one vasoactiveagent using infrared thermography, and (iv) comparing the measurementsobtained in steps (i) and (iii), wherein a vasoactive agent that resultsin an increase in the temperature is a vasodilator and a vasoactive thatresults in a decrease in the temperature is a vasoconstrictor. Inanother embodiment, a plurality of vasoactive agents may be tested inthe methods of the present invention, and the results of each test canbe compared to determine which vasoactive agent is the most effective,i.e., which vasoactive agent produces the greatest increase intemperature between step (i) amd step (iii). The baseline temperature instep (i) may be a previously obtained measurement; or alternatively ameasurement taken after the administration of the compound when a stablebaseline temperature measurement is obtained. The vasoactive agent canoptionally be administered with a nitric oxide donor. Contemplatedvasoactive agents and nitric oxide donors include all those known in theart and those described herein. The nitric oxide donors and/orvasoactive agents can be administered separately or in the form of acomposition. The compounds and compositions of the present invention canalso be administered in combination with other medications used formonitoring diseases or disorders.

[0152] Suitable vasoactive agents that can be used for the methodsdescribed herein include, but are not limited to, potassium channelactivators (such as, for example, nicorandil, pinacidil, cromakalim,minoxidil, aprilkalim, loprazolam and the like); calcium channelblockers (such as, for example, nifedipine, veraparmil, diltiazem,gallopamil, niludipine, nimodipins, nicardipine, and the like);β-blockers (such as, for example, butixamine, dichloroisoproterenol,propanolol, alprenolol, bunolol, nadolol, oxprenolol, perbutolol,pinodolol, sotalol, timolol, metoprolol, atenolol, acebutolol,bevantolol, pafenolol, tolamodol, and the like); long and short actingα-adrenergic receptor antagonists (such as, for example,phenoxybenzamide, dibenamine, doxazosin, terazosin, phentolamine,tolazoline, prozosin, trimazosin, yohimbine, moxisylyte and the like);prostaglandins (such as, for example, PGE₁, PGA₁, PGB₁, PGF₁, PGF₂,19-hydroxy-PGA¹, 19-hydroxy-PGB₁, PGE₂, PGA₂, PGB₂, prostacyclins,thromboxanes, leukotrienes, 6-keto-PGE₁ derivatives and carbacyclinderivatives, and the like); phosphodiesterase inhibitors (such as, forexample, papaverine, zaprinast, sildenafil, IC 351); adenosine, ergotalkaloids (such as, for example, ergotamine, ergotamine analogs,including, for example, acetergamine, brazergoline, bromerguride,cianergoline, delorgotrile, disulergine, ergonovine maleate, ergotaminetartrate, etisulergine, lergotrile, lysergide, mesulergine, metergoline,metergotamine, nicergoline, pergolide, propisergide, proterguride,terguride and the like); vasoactive intestinal peptides (such as, forexample, peptide histidine isoleucine, peptide histidine methionine,substance P, calcitonin gene-related peptide, neurokinin A, bradykinin,neurokinin B, and the like); dopamine agonists (such as, for example,apomorphine, bromocriptine, testosterone, cocaine, strychnine, and thelike); opioid antagonists (such as, for example, naltrexone, and thelike); endothelin antagonists (such as, for example, bosentan,sulfonamide endothelin antagonists, BQ-123, SQ 28608, and the like);thromboxane inhibitors (such as, for example, SQ 29548, BAY u3405, GR32191, YM 158, and the like), and mixtures thereof. Suitable vasoactiveagents are described more fully in the literature, such as in Goodmanand Gilman, The Pharmacological Basis of Therapeutics (9th Edition),McGraw-Hill, 1995; and the Merck Index on CD-ROM, Twelfth Edition,Version 12:1, 1996; and on STN Express, file phar and file registry.

[0153] Another embodiment of the present invention comprises methods formonitoring and measuring sexual dysfunctions and/or sexual arousal inpatients, preferably males and females, most preferably females, usinginfrared thermography resulting from the administration of at least onenitric oxide donor and/or at least one vasoactive agent. For example,the method for monitoring and measuring female sexual dysfunction maycomprise (i) measuring the baseline temperature of the gentialia usinginfrared thermography, (ii) administering to a patient at least onenitric oxide donor and/or at least one vasoactive agent, (iii) measuringthe temperature of the gentialia during and/or after administering to apatient the nitric oxide donor and/or the vasoactive agent usinginfrared thermography, and (iv) comparing the measurements obtained insteps (i) and (iii), wherein a compound that results in an increase inthe temperature between step (i) and step (iii) is a sexual enhancer anda compound that results in a decrease in the temperature between step(i) and step (iii) is a sexual inhibitor. The baseline temperature instep (i) may be a previously obtained measurement; or alternatively ameasurement taken after the administration of the compound when a stablebaseline temperature measurement is obtained. Contemplated nitric oxidedonors and vasoactive agents include all those known in the art andthose described herein. The nitric oxide donors and/or vasoactive agentscan be administered separately or in the form of a composition. Thecompounds and compositions of the present invention can also beadministered in combination with other medications used for monitoringdiseases or disorders.

[0154] Yet another embodiment of present invention comprises methods formonitoring and diagnosing diseases and disorders resulting fromvasodilation and changes in blood flow using infrared thermography. Thediseases and disorders resulting from changes in vasodilation and bloodflow include, or example, Raynaud's syndrome, inflammation,hypertension, gastrointestinal disorders and central nervous systemdisorders. For example, the method for monitoring and diagnosingdiseases and disorders resulting from vasodilation and changes in bloodflow may comprise (i) measuring the baseline temperature of an area ofinterest using infrared thermography, (ii) administering to a patient atleast one nitric oxide donor and/or at least one vasoactive agent, (iii)measuring the temperature of the area of interest during and/or afteradministering to a patient the at least one nitric oxide donor and/or atleast one vasoactive agent using infrared thermography, and (iv)comparing the measurements obtained in steps (i) and (iii), wherein acompound that results in an increase in the temperature between step (i)and step (iii) will be useful for treating the diseases and disordersdescribed herein. The baseline temperature in step (i) may be apreviously obtained measurement; or alternatively a measurement takenany time after the administration of the at least one nitric oxide donorand/or at least one vasoactive agent when a stable baseline temperaturemeasurement is obtained. Contemplated nitric oxide donors and vasoactiveagents include all those known in the art and those described herein.The nitric oxide donors and/or vasoactive agents can be administeredseparately or in the form of a composition. The compounds andcompositions of the present invention can also be administered incombination with other medications used for monitoring diseases ordisorders.

[0155] Yet another embodiment of present invention comprises methods foridentifying, characterizing, ranking and selecting compounds for thetreatment of a disease and disorder resulting from vasodilation andchanges in blood flow using infrared thermography. For example, themethod for identifying, characterizing, ranking and selecting compoundsfor the treatment of a disease and disorder resulting from vasodilationand changes in blood flow may comprise (i) measuring the baselinetemperature of an area of interest using infrared thermography, (ii)administering to a patient a test compound, (iii) measuring thetemperature of the area of interest during and/or after administering toa patient the test compound using infrared thermography, and (iv)comparing the measurements obtained in steps (i) and (iii), wherein acompound that results in an increase in the temperature between step (i)and step (iii) is effective for the treatment of the disease or disorderand a compound that produces no temperature change or results in adecrease in the temperature between step (i) and step (iii) is noteffective for the treatment of the disease or disorder. The greater theincrease in temperature of the area of interest between step (i) andstep (iii) the more potent the compound. In another embodiment, aplurality of test compounds can be tested in the methods of the presentinvention, and the results from each test can be compared to determinewhich test compound is the most effective, i.e., which test compoundproduces the greatest increase in temperature between step (i) and step(iii). In some instances it may be desirable to use the methods of thepresent invention to find compounds which produce the greatest decreasein temperature between step (i) and step (ii), or which do not produceany change in temperature between step (i) and step (iii). The baselinetemperature in step (i) may be a previously obtained measurement; oralternatively a measurement taken any time after the administration ofthe test compound when a stable baseline temperature measurement isobtained.

[0156] Yet another embodiment of the present invention providescompositions comprising at least one S-nitrosothiol compound and atleast one penetration enhancers that may be used to diagnose, monitorand/or treat female sexual dysfunctions. The S-nitrosothiol compound maypreferably be S-nitrosoglutathione. The penetration enhancer, maypreferably be a glyceride, such as, MIGLYOL®, and/or a polyglcolyzedglyceride, such as, LABROSOL® and/or LABROFIL®, or mixtures thereof.These compositions may further comprise at least one vascoactive agentand/or at least one nitric oxide donor, or mixtures thereof.

[0157] In a particular embodiment, the glyceride penetration enhancerMIGLYOL® is MIGLYLOL® 812N obtained from Condea Vista Company, Houston,Tex. MIGLYOL® 812N is a mixture of caprylic triglycerides and caprictriglycerides. It can also contain decanoly triglycerides, octanoyltriglycerides and C₈-C₁₂ triglycerides.

[0158] The polyglycolyzed glyceride may be saturated or unsaturated andmay include ethoxylated glycerides and polyethylene glycol esters. In aparticular embodiment, the saturated polyglycolyzed glyceride is aglyceryl caprylate/caprate and PEG-8 (polyethylene glycol)caprylate/caprate complex known as LABRASOL® (Gattefosse Corp., NewYork). Suitable unsaturated polyglycolyzed glycerides are apricot kerneloil PEG-6 complex (LABRAFIL® M-1944 CS), almond oil PEG-6 complex(LABRAFIL® M-1966 CS), peanut oil PEG-6 complex (LABRAFIL® M-1969 CS),olive oil PEG-6 complex (LABRAFIL® M-1980 CS) and corn oil PEG-6 complex(LABRAFIL® M-2125 CS), all available from Gattefosse Corp., New York.Suitable ethoxylated glyceride, include, but are not limited to, C₈-C₁₀carbon chain, for example glyceryl caprylate/caprate PEG-4 complex.

[0159] When administered in vivo, the nitric oxide donors and/orvasoactive agents of the present invention may be administered withpharmaceutically acceptable carriers and in dosages described herein.When the nitric oxide donors and/or vasoactive agents of the presentinvention are administered as a mixture of at least one nitric oxidedonor and at least one vasoactive agent, they can also be used incombination with one or more additional compounds (e.g., therapeuticagents used to treat, diagnose and monitor the disease and disorder).When administered separately, the nitric oxide donor(s) and/orvasoactive agent can be administered simultaneously with, subsequentlyto, or prior to administration of the other additional compound(s) totreat or monitor the diseases described herein.

[0160] The compounds and compositions of the present invention can beadministered by any available and effective delivery system including,but not limited to, orally, bucally, parenterally, by inhalation spray(oral or nasal), by topical application, by injection into the corpuscavernosum tissue, by transurethral drug delivery, vaginally, orrectally (e.g., by the use of suppositories) in dosage unit formulationscontaining conventional nontoxic pharmaceutically acceptable carriers,adjuvants, and vehicles, as desired. Parenteral includes subcutaneousinjections, intravenous injections, intramuscular injections,intrasternal injections, and infusion techniques. Parenteral alsoincludes injection into the corpus cavernosum tissue, which can beconducted using any effective injection system including, but notlimited to, conventional syringe-and-needle systems or needlelessinjection devices.

[0161] Solid dosage forms for oral administration can include capsules,tablets, effervescent tablets, chewable tablets, pills, powders,effervescent powders, sachets, granules and gels. In such solid dosageforms, the active compounds can be admixed with at least one inertdiluent such as sucrose, lactose or starch. Such dosage forms can alsocomprise, as in normal practice, additional substances other than inertdiluents, e.g., lubricating agents such as magnesium stearate. In thecase of capsules, tablets, effervescent tablets, and pills, the dosageforms can also comprise buffering agents. Soft gelatin capsules can beprepared to contain a mixture of the active compounds or compositions ofthe present invention and vegetable oil. Hard gelatin capsules cancontain granules of the active compound in combination with a solid,pulverulent carrier such as lactose, saccharose, sorbitol, mannitol,potato starch, corn starch, amylopectin, cellulose derivatives ofgelatin. Tablets and pills can be prepared with enteric coatings.

[0162] Liquid dosage forms for oral administration can includepharmaceutically acceptable emulsions, solutions, suspensions, syrups,and elixirs containing inert diluents commonly used in the art, such aswater. Such compositions can also comprise adjuvants, such as wettingagents, emulsifying and suspending agents, and sweetening, flavoring,and perfuming agents.

[0163] Suppositories for vaginal or rectal administration of thecompounds and compositions of the invention can be prepared by mixingthe compounds or compositions with a suitable nonirritating excipientsuch as cocoa butter and polyethylene glycols which are solid at roomtemperature but liquid at body temperature, such that they will melt andrelease the drug.

[0164] Injectable preparations, for example, sterile injectable aqueousor oleaginous suspensions can be formulated according to the known artusing suitable dispersing agents, wetting agents and/or suspendingagents. The sterile injectable preparation can also be a sterileinjectable solution or suspension in a nontoxic parenterally acceptablediluent or solvent, for example, as a solution in 1,3-butanediol. Amongthe acceptable vehicles and solvents that can be used are water,Ringer's solution, and isotonic sodium chloride solution. Sterile fixedoils are also conventionally used as a solvent or suspending medium.

[0165] Topical administration, which is well known to one skilled in theart, involves the delivery of pharmaceutical agents via percutaneouspassage of the drug into the systemic circulation of the patient.Topical administration includes vaginal administration, vulvaladministration, penile administration and rectal administration. Topicaladministration can also involve transdermal patches or iontophoresisdevices. Other components can be incorporated into the transdermalpatches as well. For example, compositions and/or transdermal patchescan be formulated with one or more preservatives or bacteriostaticagents including, but not limited to, methyl hydroxybenzoate, propylhydroxybenzoate, chlorocresol, benzalkonium chloride, and the like.

[0166] Dosage forms for topical administration of the compounds andcompositions of the present invention preferably include creams, sprays,lotions, gels, ointments, emulsions, coatings for condoms, liposomes,foams, and the like. Administration of the cream, spray, ointment,lotion, gel, emulsion, coating, liposome, or foam can be accompanied bythe use of an applicator or by transurethral drug delivery using asyringe with or without a needle or penile insert or device, or byclitoral, vulval or vaginal delivery, and is within the skill of theart. Alternatively, the compositions may be contained within a vaginalring, tampon, suppository, sponge, pillow, puff, or osmotic pump system;these platforms are useful solely for vaginal delivery. Typically alubricant and/or a local anesthetic for desensitization can also beincluded in the formulation or provided for use as needed. Lubricantsinclude, for example, K-Y jelly (available from Johnson & Johnson) or alidocaine jelly, such as XYLOCAINE® 2% jelly (available from AstraPharmaceutical Products). Local anesthetics include, for example,novocaine, procaine, tetracaine, benzocaine and the like.

[0167] Ointments are semisolid preparations that are typically based onpetrolatum or other petroleum derivatives. The specific ointment base tobe used, as will be appreciated by those skilled in the art, is one thatwill provide for optimum drug delivery. An ointment base should beinert, stable, nonirritating and nonsensitizing. Ointment bases may begrouped in four classes: oleaginous bases; emulsifiable bases; emulsionbases; and water-soluble bases. Oleaginous ointment bases include, forexample, vegetable oils, fats obtained from animals, semisolidhydrocarbons obtained from petroleum, and the like. Emulsifiableointment bases, also known as absorbent ointment bases, contain littleor no welter and include, for example, hydroxystearin sulfate, anhydrouslanolin, hydrophilic petrolatum, and the like. Emulsion ointment basesare either water-in-oil (W/O) emulsions or oil-in-water (O/W) emulsions,and include, for example, cetyl alcohol, glyceryl monostearate, lanolinand stearic acid, and the like. In a particular embodiment,water-soluble ointment bases are preferred and are prepared frompolyethylene glycols of varying molecular weight, and can be determinedby standard techniques as described in Remington: The Science andPractice of Pharmacy.

[0168] Lotions are preparations that may be applied without friction,and are typically liquid or semiliquid preparations in which solidparticles, including the active agent, are present in a water or alcoholbase. Lotions are usually suspensions of solids, and in a particularembodiment, may comprise a liquid oily emulsion of the oil-in-watertype. It is generally necessary that the insoluble matter in a lotion befinely divided. Lotions will typically contain suspending agents toproduce better dispersions as well as compounds useful for localizingthe active agent in contact with the skin, such as, for example,methylcellulose, sodium carboxymethyl-cellulose, and the like.

[0169] Emulsion formulations are generally formed from a dispersed phase(for example., a pharmacologically active agent), a dispersion mediumand an emulsifing agent. If desired, emulsion stabilizers can beincluded in the formulation as well. A number of pharmaceutically usefulemulsions are known in the art, including, for example, oil-in-water(o/w) formulations, water-in-oil (w/o) formulations and multipleemulsions such as w/o/w or o/w/o formulations. Emulsifying agentssuitable for use in such formulations include, but are not limited to,TWEEN 60®, SPAN 80®, cetostearyl alcohol, myristyl alcohol, glycerylmonostearate, sodium lauryl sulfate, and the like.

[0170] Creams are, as known in the art, viscous liquid or semisolidemulsions, either oil-in-water or water-in-oil. Cream bases arewater-washable, and contain an oil phase, an emulsifier and an aqueousphase. The oil phase, also sometimes called the “internal” phase, isgenerally comprised of petrolatum and a fatty alcohol such as, cetylalcohol, stearyl alcohol, and the like; the aqueous phase usually,although not necessarily, exceeds the oil phase in volume, and generallycontains a humectant. The emulsifier in a cream formulation is generallya nonionic, anionic, cationic or amphoteric surfactant.

[0171] The ointments, lotions, emulsions and creams are formed bydispersing finely divided or dissolved the nitric oxide donor(s) and/orvasoactive agent(s) uniformly throughout the vehicle or base usingconventional techniques, typically by levigating the compound with asmall quantity of the base to form a concentrate which is then dilutedgeometrically with further base. Alternatively, a mechanical mixer maybe used. Creams, lotions and emulsions are formed by way of a two-phaseheat system, wherein oil-phase ingredients are combined under heat toprovide a liquified, uniform system. The aqueous-phase ingredients areseparately combined using heat. The oil and aqueous phases are thenadded together with constant agitation and allowed to cool. At thispoint, concentrated agents may be added as a slurry. Volatile oraromatic materials can be added after the emulsion has sufficientlycooled. Preparation of such pharmaceutical compositions is within thegeneral skill of the art. See, e.g., Remington's PharmaceuticalSciences, 18th Ed. (Easton, Pa.: Mack Publishing Company, 1990).

[0172] The vasoactive agents can also be incorporated into gelformulations using known techniques. Two-phase gel systems generallycomprise a suspension or network of small, discrete particlesinterpenetrated by a liquid to provide a dispersed phase and a liquidphase. Single-phase gel systems are formed by distributing organicmacromolecules uniformly throughout a liquid such that there are noapparent boundaries between the dispersed and liquid phases. Suitablegelling agents for use herein include synthetic macromolecules, such as,CARBOMERS®, polyvinyl alcohols, and polyoxyethylene-polyoxypropylenecopolymers, and the like; gums such as, tragacanth, as well as sodiumalginate, gelatin, methylcellulose, sodium carboxymethylcellulose,methylhydroxyethyl cellulose, hydroxyethyl cellulose, and the like. Inorder to prepare a uniform gel, dispersing agents such as alcohol orglycerin may be added, or the gelling agent may be dispersed bytrituration, mechanical mixing or stirring, or combinations thereof.

[0173] The compounds and compositions of the present invention willtypically be administered in a pharmaceutical composition containing oneor more carriers or excipients, i.e., pharmaceutically acceptableorganic or inorganic carrier substances suitable for parenteralapplication which do not deleteriously react with the active compounds.Examples of pharmaceutically acceptable carriers include, for example,water, salt solutions, alcohol, silicone, waxes, petroleum jelly,vegetable oils, polyethylene glycols, propylene glycol, liposomes,sugars, gelatin, lactose, amylose, magnesium stearate, talc,surfactants, silicic acid, viscous paraffin, perfume oil, fatty acidmonoglycerides and diglycerides, petroethral fatty acid esters,hydroxymethyl-cellulose, polyvinylpyrrolidone, and the like.

[0174] The pharmaceutical preparations can be sterilized and if desired,mixed with auxiliary agents which do not deleteriously react with theactive compounds, e.g., lubricants, preservatives, stabilizers, wettingagents, emulsifiers, salts for influencing osmotic pressure, buffers,colorings, flavoring and/or aromatic substances, and the like. Forparenteral application, particularly suitable vehicles consist ofsolutions, preferably oily or aqueous solutions, as well as suspensions,emulsions, or implants. Aqueous suspensions may contain substances whichincrease the viscosity of the suspension and include, for example,sodium carboxymethyl cellulose, sorbitol and/or dextran. Optionally, thesuspension may also contain stabilizers.

[0175] The pharmaceutical compositions may also include a detergent inthe formulation, in an amount effective to increase solubility of thenitric oxide donor and/or vasoactive agent in the vehicle andbioavailability of the agent following administration. The detergentwill typically be a nonionic, anionic, cationic or amphotericsurfactant. The surfactant is selected such that local irritation at thesite of administration is avoided. Surfactants include, for example,TERGITOL.® and TRITON® surfactants (Union Carbide Chemicals andPlastics, Danbury, Conn. polyoxyethylene sorbitan fatty acid esters,e.g., TWEEN® surfactants (Atlas Chemical Industries, Wilmington, Del.),such as, for example, polyoxyethylene 20 sorbitan monolaurate (TWEEN®20), polyoxyethylene (4) sorbitan monolaurate (TWEEN® 21),polyoxyethylene 20 sorbitan monopalmitate (TWEEN® 40), polyoxyethylene20 sorbitan monooleate (TWEEN®D 80, and the like; polyoxyethylene 4lauryl ether (BRIJ® 30), polyoxyethylene 23 lauryl ether (BRIJ 35),polyoxyethylene 10 oleyl ether (BRIJ® 97); polyoxyethylene glycolesters, such as, for example, poloxyethylene 8 stearate (MYRJ® 45),poloxyethylene 40 stearate (MYRJ® 52) polyoxyethylene alkyl ethers, andthe like; or mixtures thereof.

[0176] The pharmaceutical preparation may also include one or morepermeation enhancers. Permeation enhancers include those generallyuseful in conjunction with topical, transdermal or transmucosal drugdelivery. Permeation enhancers include, for example, dimethylsulfoxide(DMSO), dimethyl formamide (DMF), N,N-dimethylacetamide (DMA),decylmethylsulfoxide (C₁₀ MSO), polyethylene glycol monolaurate (PEGML),polyethyleneglycol, glycerol monolaurate, lecithin, the 1-substitutedazacycloheptan-2-ones, such as, 1-n-dodecylcyclazacycloheptan-2-one(available under the trademark AZONE® from Nelson Research & DevelopmentCo., Irvine, Calif.), lower alkanols (e.g., ethanol), C₆ toC₂₀-hydrocarbyl substituted 1,3-dioxane, C₆ to C₂₀-hydrocarbylsubstituted 1,3-dioxolane and C₆ to C₂₀-hydrocarbyl substituted acetal,such as, SEPA® (available from Macrochem Co., Lexington, Mass.),alkonates, such as, alkyl-2-(N,N-disubstituted amino)-alkonate ester,N,N-disubstituted amino)-alkanol alkanoate, and the like, glycerides,such as mono, di and triglycerides and mixtures thereof, such as forexample MIGLYOL® (Condea Vista Company, Houston, Tex.) and the like;polyglycolyzed glycerides, such as, for example, LABRASOL® andLABRAFIL®, and the like; and surfactants as discussed above, including,for example, TERGITOL.® and TRITON® surfactants, NONOXYNOL-9® andTWEEN-80®. In particular embodiments the penetration enhancers may beMIGLYOL®, LABRASOL® or LABRAFIL®, including mixtures thereof.

[0177] In some cases, the formulations may include one or more compoundseffective to inhibit enzymes present in the vaginal or vulvar areaswhich could degrade or metabolize the pharmacologically active agent.For example, with a prostaglandin as the vasoactive agent, it may bepreferred to include an effective inhibiting amount of a compoundeffective to inhibit prostaglandin-degrading enzymes. Such compoundsinclude, for example, fatty acids, fatty acid esters, and NADinhibitors.

[0178] Various delivery systems are known and can be used to administerthe compounds or compositions of the present invention, including, forexample, encapsulation in liposomes, microbubbles, emulsions,microparticles, microcapsules and the like. The required dosage can beadministered as a single unit or in a sustained release form.

[0179] The bioavailabilty of the compositions can be enhanced bymicronization of the formulations using conventional techniques such asgrinding, milling, spray drying and the like in the presence of suitableexcipients or agents such as phospholipids or surfactants. Thebioavailability and absorption of the nitric oxide donor and/orvasoactive agent can be increased by the addition of tablettingexcipients, such as, for example β-cyclodextrin, a β-cyclodextrinderivative, such as for example, hydroxypropyl-β-cyclodextrin (HPBCD),and the like. Inclusion complexes are complexes formed by interaction ofmacrocyclic compounds containing an intramolecular cavity of moleculardimensions with the smaller, pharmacologically active agent. Preferredinclusion complexes are formed from α-, β- and γ-cyclodextrins, or fromclathrates, in which the “host” molecules form a crystal latticecontaining spaces in which “guest” molecules (i.e., in this case, thenitric oxide donor and/or vasoactive agent) will fit. See, e.g., Hagan,Clathrate Inclusion Compounds (New York: Reinhold, 1962).

[0180] Liposomes are microscopic vesicles having a lipid wall comprisinga lipid bilayer, and can be used as drug delivery systems as well.Generally, liposome formulations are preferred for poorly soluble orinsoluble pharmaceutical agents. Liposomal preparations for use in thepressent invention include cationic (positively charged), anionic(negatively charged) and neutral preparations. Cationic liposomes arereadily available. For example, N(1-2,3-dioleyloxy)propyl)-N,N,N-triethylammonium (DOTMA) liposomes are available under thetradename LEPOFECTIN® (GIBCO BRL, Grand Island, N.Y.). Similarly,anionic and neutral liposomes are readily available as well, from AvantiPolar Lipids (Birmingham, Ala.), or can be easily prepared using readilyavailable materials. Such materials include phosphatidyl choline,cholesterol, phosphatidyl ethanolamine, dioleoylphosphatidyl choline(DOPC), dioleoylphosphatidyl glycerol (DOPG), dioleoylphoshatidylethanolamine (DOPE), and the like. These materials can also be mixedwith DOTMA in appropriate ratios. Methods for making liposomes usingthese materials are well known in the art. See Remington'sPharmaceutical Sciences, supra.

[0181] The release of the nitric oxide donor and/or vasoactive agent canbe controlled by dissolution (bioerosion) of a polymer using eitherencapsulated dissolution control or matrix dissolution control. Inencapsulated dissolution control, the vasoactive agent is coated with amembrane of slowly dissolving polymeric or wax materials. When theencapsulating membrane has dissolved, the agent core is available forimmediate release and adsorption across the epithelial or mucosalsurfaces of the vagina or vulvar area. Bioerodible coating materials maybe selected from a variety of natural and synthetic polymers, dependingon the agent to be coated and the desired release characteristics.Exemplary coating materials include gelatins, carnauba wax, shellacs,ethylcellulose, cellulose acetate phthalate, cellulose acetate butyrate,and the like. Release of the compound is controlled by adjusting thethickness and dissolution rate of the polymeric membrane. A uniformsustained release can be attained by compressing a population ofparticles of the agent with varying membrane thickness (e.g., varyingerosion times) into a tablet form for a single administration.

[0182] In matrix dissolution control, the nitric oxide donor and/orvasoactive agent is dissolved or dispersed within a matrix of, such as,for example, an erodible wax. The compound is released for adsorptionacross the epithelial or mucosal surfaces of the vagina or vulvar areaas the matrix bioerodes. The rate of compound availability is generallycontrolled by the rate of penetration of the dissolution media (i.e.,vaginal fluids) into the matrix, wherein the rate of penetration isdependent on the porosity of the matrix material. Bioerodible matrixdissolution delivery systems can be prepared by compressing the nitricoxide donor and/or vasoactive agent with a slowly soluble polymercarrier into a tablet or suppository form. There are several methods ofpreparing drug/wax particles including congealing and aqueous dispersiontechniques. In congealing methods, the vasodilating agent is combinedwith a wax material and either spray-congealed, or congealed and thenscreened. For an aqueous dispersion, the vasodilating agent/waxcombination is sprayed or placed in water and the resulting particlescollected. Matrix dosage formulations can be formed by compaction orcompression of a mixture of vasodilating agent, polymer and excipients.

[0183] In an alternative embodiment, the compositions of the presentinvention may be administered as biodegradable adhesive film or sheetwhich adhere to the vulvar area. Such drug delivery systems aregenerally composed of a biodegradable adhesive polymer based on apolyurethane, a poly(lactic acid), a poly(glycolic acid), a poly(orthoester), a polyanhydride, a polyphosphazene, or a mixture or copolymerthereof. Preferred biodegradable adhesive polymers include, for example,polyurethanes and block copolyurethanes containing peptide linkages,simple mixtures of polyurethanes and polylactides, and copolymers ofacrylates and mono- or disaccharide residues.

[0184] The compounds and compositions of the present invention can beformulated as pharmaceutically acceptable salts. Pharmaceuticallyacceptable salts include, for example, alkali metal salts and additionsalts of free acids or free bases. The nature of the salt is notcritical, provided that it is pharmaceutically acceptable. Suitablepharmaceutically acceptable acid addition salts may be prepared from aninorganic acid or from an organic acid. Examples of such inorganic acidsinclude, but are not limited to, hydrochloric, hydrobromic, hydroiodic,nitric (nitrate salt), nitrous (nitrite salt), carbonic, sulfuric andphosphoric acid and the like. Appropriate organic acids include, but arenot limited to, aliphatic, cycloaliphatic, aromatic, heterocyclic,carboxylic and sulfonic classes of organic acids, such as, for example,formic, acetic, propionic, succinic, glycolic, gluconic, lactic, malic,tartaric, citric, ascorbic, glucuronic, maleic, fumaric, pyruvic,aspartic, glutamic, benzoic, anthranilic, mesylic, salicylic,p-hydroxybenzoic, phenylacetic, mandelic, embonic (pamoic),methanesulfonic, ethanesulfonic, benzenesulfonic, pantothenic,toluenesulfonic, 2-hydroxyethanesulfonic, sulfanilic, stearic, algenic,β-hydroxybutyric, cyclohexylaminosulfonic, galactaric and galacturonicacid and the like. Suitable pharmaceutically acceptable base additionsalts include, but are not limited to, metallic salts made fromaluminum, calcium, lithium, magnesium, potassium, sodium and zinc ororganic salts made from primary, secondary and tertiary amines, cyclicamines, N,N′-dibenzylethylenediamine, chloroprocaine, choline,diethanolamine, ethylenediamine, meglumine (N-methylglucamine) andprocaine and the like. All of these salts may be prepared byconventional means from the corresponding compound by reacting, forexample, the appropriate acid or base with the compound.

[0185] “Therapeutically effective amount” refers to the amount of thenitric oxide donor and/or vasoactive agent which is effective to achieveits intended purpose. In preferred embodiments of the methods describedherein, the nitric oxide donor and/or vasoactive agents are administeredin a therapeutically effective amount. While individual patient needsmay vary, determination of optimal ranges for effective amounts of eachnitric oxide donor is within the skill of the art. Generally the dosageregimen for monitoring and idagnosing a condition with the compoundsand/or compositions of this invention is selected in accordance with avariety of factors, including the type, age, weight, sex, diet andmedical condition of the patient, the severity of the dysfunction, theroute of administration, pharmacological considerations such as theactivity, efficacy, pharmacokinetic and toxicology profiles of theparticular compound used, whether a drug delivery system is used, andwhether the compound is administered as part of a drug combination andcan be adjusted by one skilled in the art. Thus, the dosage regimenactually employed may vary from the preferred dosage regimen set forthherein.

[0186] The amount of a given nitric oxide donor and/or vasoactive agentwhich will be effective in monitoring and diagnosing a particulardysfunction or condition will depend on the nature of the dysfunction orcondition, and can be determined by standard clinical techniques,including reference to Goodman and Gilman, supra; The Physician's DeskReference, supra; Medical Economics Company, Inc., Oradell, N.J., 1995;and Drug Facts and Comparisons, Inc., St. Louis, Mo., 1993. The precisedose to be used in the formulation will also depend on the route ofadministration, and the seriousness of the dysfunction or disorder, andshould be decided by the physician and the patient's circumstances.Effective doses may be extrapolated from dose-response curves derivedfrom in vitro or animal model test systems and are in the same ranges orless than as described for the commercially available compounds in thePhysician's Desk Reference, supra.

[0187] In particular embodiments the methods of administration of thenitric oxide donors and/or vasoactive agents for monitoring, diagnosingand treating male sexual dysfunction are by oral administration, bytopical application, by injection into the corpus cavernosum, bytransurethral administration or by the use of suppositories. Thepreferred methods of administration for monitoring, diagnosing andtreating female sexual dysfunction are by oral administration, topicalapplication or by the use of suppositories. The most preferred mode ofadministration for female sexual dysfunction is topical application,preferably as an ointment, a cream, a gel, an emulsion, a spray or alotion. These compositions may contain at least one penetration enhancerto increase the premeability of the nitric oxide donor and/or vasoactiveagent across the membrane.

[0188] The doses of nitric oxide donors for monitoring and diagnosingsexual dysfunction in the pharmaceutical composition can be in amountsof about 0.001 mg to about 30 g and the actual amount administered willbe dependent on the specific nitric oxide donor compound. For example,when L-arginine is the nitric oxide donor, L-arginine can beadministered orally in an amount of about 0.25 grams to about 10 grams(equivalent to about 0.5 grams to about 20 grams of L-arginineglutamate), preferably about 2 grams to about 4 grams (equivalent toabout 4 grams to about 8 grams of L-arginine glutamate); more preferablyabout 2.5 grams to about 3.5 grams (equivalent to about 5 grams to about7 grams of L-arginine glutamate); most preferably about 3 grams(equivalent to 6 grams of L-arginine glutamate).

[0189] The α-antagonist, such as phentolamine, can be administered inamounts of about 3.7 mg to about 90 mg (equivalent to about 5 mg toabout 120 mg phentolamine mesylate), preferably about 22 mg to about 37mg (equivalent to about 30 mg to about 50 mg phentolamine mesylate),more preferably about 26 mg to about 34 mg (equivalent to about 35 mg toabout 45 mg phentolamine mesylate), even more preferably about 28 mg toabout 31 mg (equivalent to about 38 mg to about 42 mg phentolaminemesylate), most preferably about 30 mg (equivalent to about 40 mgphentolamine mesylate).

[0190] The α-antagonist, such as yohimbine, can be administered in anamount of about 1.0 mg to about 18.0 mg (equivalent to about 1.1 mg toabout 19.8 mg yohimbine hydrochloride), preferably about 4.5 mg to about6.4 mg, (equivalent to about 5.0 mg to about 7.0 mg yohimbinehydrochloride), more preferably about 5.0 mg to about 6.0 mg,(equivalent to about 5.5 mg to about 6.5 mg yohimbine hydrochloride),most preferably about 5.5 mg (equivalent to about 6.0 mg yohimbinehydrochloride). The yohimbine can also be administered in the form ofits pharmaceutical salt, yohimbine tartarate, or yohimbe bark powder orextract that has been standardized to deliver up to about 18 mg ofyohimbine.

[0191] The present invention also provides pharmaceutical kitscomprising one or more containers filled with one or more of theingredients of the pharmaceutical compounds and/or compositions of thepresent invention, including, one or more NO donors, and one or morevasoactive agents. Such kits can also include, for example, othercompounds and/or compositions (e.g., permeation enhancers, lubricants,and the like), a device(s) for administering the compounds and/orcompositions, and written instructions in a form prescribed by agovernmental agency regulating the manufacture, use or sale ofpharmaceuticals or biological products, which instructions can alsoreflects approval by the agency of manufacture, use or sale for humanadministration.

EXAMPLES

[0192] The following non-limiting examples are for purposes ofillustration only and are not intended to limit the scope of theinvention or claims.

Example 14-Aza-4-(2-methyl-2-(nitrosothio)propyl)tricyclo(5.2.1.0<2,6>)dec-8-ene-3,5-dione

[0193] 1a.4-Aza4-(2-methyl-2-sulfanylpropyl)tricyclo(5.2.1.0<2,6>)dec-8-ene-3,5-dione

[0194] A suspension of 1-amino-2-methylpropane-2-thiol hydrochloride(6.72 g, 47.4 mmol) in ethyl acetate (200 mL) was shaken with potassiumhydroxide solution (16 M, 3.6 mL, 57.0 mmol). The ethyl acetate solutionwas separated, dried with sodium sulfate, filtered, and concentrated togive 1-amino-2-methylpropane-2-thiol (2.70 g, 25.7 mmol, 54%). The thiolwas dissolved in acetic acid (25 mL) andcis-5-norbornene-endo-2,3-dicarboxylic anhydride (4.17 g, 25.4 mmol) wasadded. The reaction was stirred at 100° C. for 1 hour and allowed tostand at room temperature over the weekend. The crystals which formedwere collected by filtration, washed with acetic acid (4 mL) and a smallvolume of methanol, and then dried in vacuo to give the title compound(2.22 g, 35%). The filtrate was concentrated, treated with toluene andconcentrated (repeat four times). The residue dissolved indichloromethane and filtered through silica gel to give additionalproduct (2.47 g) contaminated with a littlecis-5-norbornene-endo-2,3-dicarboxylic anhydride. ¹H NMR (CDCl₃) δ 6.16(s, 2H), 3.52 (s, 2H), 3.42 (s, 2H), 3.32 (s, 2H), 1.86 (s, 1H), 1.76(d, J=8.77 Hz, 1H), 1.57 (d, J=8.77 Hz, 1H), 1.30 (s, 6H). ¹³C NMR(CDCl₃) δ 177.9, 134.8, 52.5, 51.0, 45.8, 45.24, 45.0, 30.9. LRMS(APIMS) m/z 252 (MH⁺).

[0195] 1b.4-Aza-4-(2-methyl-2-(nitrosothio)propyl)tricyclo(5.2.1.0<2,6>)dec-8-ene-3,5-dione

[0196] To a solution of Example 1a (793 mg, 3.156 mmol) indichloromethane (23 mL) was added tert-butyl nitrite (750 μL, 650 mg,6.31 mmol) and the solution was stirred at room temperature for 1 hourin the dark. The reaction mixture was concentrated and the residuechromatographed (ethyl acetate:hexane 2:3) to give the title compound(768.7 mg, 2.738 mmol, 87%). ¹H NMR (CDCl₃) δ 6.12 (s, 2H), 4.10 (s,2H), 3.41 (s, 2H), 3.30 (s, 2H), 1.82 (s, 6H), 1.75 (d, J=8.8 Hz, 1H),1.57 (d, J=8.8 Hz, 1H). ¹³C NMR (CDCl₃) δ 177.7, 134.7, 56.7, 52.4,48.0, 47.0, 46.0, 45.8, 45.0, 27.5. LRMS (APIMS) nz/z 298 (M⁺+NH₄).99.3% purity by HPLC analysis (Column: Water μBondpack C18; Size: 3.9mm×150 mm; Solvent A: acetonitrile/0.1% TFA; Solvent B: water/0.1% TFA;Flow rate: 1.0 mL/min; Program: 20% A to 95% A over 20 min.; Detection:254 nm; Sample: 4.3 mg/mL; Injection volume: 10 μL).

Example 2 4-(1-Methyl-1-(nitrosothio)ethyl)-1,3-oxazolidin-2-one

[0197] 2a.2-Amino-3-methyl-3-((2,4,6-trimethoxyphenyl)methylthio)butanoic acid

[0198] A suspension of 2-amino-3-methyl-3-sulfanylbutanoic acid(D-penicillamine) (5.0 g, 34 mmol) in CH₂Cl₂ (150 mL) was cooled to 0°C. Trifluoroacetic acid (54 mL, 703 mmol) was added dropwise over aperiod of 5 minutes. Then 2,4,6-trimethoxybenzyl alcohol (6.64 g, 34mmol) in CH₂Cl₂ (137 mL) was added dropwise at 0° C. with stirring.Stirring was continued for 1 hour at 0° C. and 2 hours at roomtemperature. The solvent was removed in vacuo and the residue was driedunder high vacuum for 3 hours. The crude red solid was recrystallizedfrom 1:1:1 CH₂Cl₂/MeOH/EtOAc to give the title compound as a white solid(10.5 g, 95%). ¹H NMR (300 MHz, CDCl₃) δ 6.10 (s, 2H), 3.84 (s, 6H),3.76 (s, 3H), 3.40-4.10 (m, 3H), 1.69 (s, 3H), 1.23 (s, 3H). LRMS (EI)m/z 330 (MH⁺).

[0199] 2b.2-Amino-3-methyl-3-((2,4,6-trimethoxyphenyl)methylthio)butan-1-ol

[0200] To a stirred solution of Example 2a (10.5 g, 32 mmol) in THF (80mL) was added dropwise lithium aluminum hydride (1 M in THF, 64 mL, 64mmol) at 0° C. under nitrogen. The resulting solution was stirred at 0°C. for 1 hour and then at room temperature for 2 hours. The excessreducing agent was destroyed carefully by portionwise addition ofNa₂SO₄.10H₂O at 0° C. The granular white precipitate was filtered andwashed with 30% methanol in CH₂Cl₂. The combined filtrates were driedover Na₂SO₄, filtered and evaporated to give the title compound as ayellow oil (7.6 g, 76%) which was used for the next step without furtherpurification. ¹H NMR (300 MHz, CDCl₃) δ 6.10 (s, 2H), 3.85 (s, 6H), 3.81(s, 3H), 3.74 (s, 2H), 3.60-3.80 (m, 2H), 3.37-3.43 (m, 1H), 2.93-2.98(m, 1H), 1.45 (s, 3H), 1.30 (s, 3H). LRMS (EI) m/z 316 (MH⁺).

[0201] 2c.4-{1-Methyl-1-((2,4,6-trimethoxyphenyl)methylthio)ethyl}-1,3-oxazolidin-2-one

[0202] A mixture of K₂CO₃ (0.33 g, 2.4 mmol), diethylcarbonate (50 mL)and the product of Example 2b (7.6 g, 24 mmol) was heated at 100° C. for24 hours. The solvent was evaporated and the resultant light brownslurry was cooled to room temperature, diluted with CH₂Cl₂ and filteredto remove the K₂CO₃. The filtrate was evaporated and the residue waschromatographed on silica gel eluting with 1:1 EtOAc:Hex to give thetitle compound as a viscous yellow oil (2.6 g, 32%). ¹H NMR (300 MHz,CDCl₃) δ 6.13 (s, 2H), 6.07 (bs, 1H), 4.30-4.40 (m, 1H), 4.25-4.28 (m,1H), 4.03-4.08 (m, 1H), 3.86 (s, 6H), 3.83 (s, 2H), 3.81 (s, 3H), 1.32(s, 3H), 1.27 (s, 3H). ¹³C NMR (75 MHz, CDCl₃) δ 160.7, 159.5, 158.7,106.3, 90.9, 66.5, 59.5, 56.0, 55.5, 47.1, 23.8, 22.3, 20.3. LRMS (EI)m/z 342 (MH⁺), 359 (MNH₄ ⁺), 364 (MN⁺).

[0203] 2d. 4-(1-Methyl-1-sulfanylethyl)-1,3-oxazolidin-2-one

[0204] The product of Example 2c (2.5 g, 7.3 mmol) was treated withwater (2.9 mL), phenol (2.9 g), anisole (2.9 mL) and finallytrifluoroacetic acid (36 mL). The resultant solution was stirred at roomtemperature for 1 hour and the solvent was evaporated to give a yellowoil. The yellow oil was dissolved in CH₂Cl₂, washed with saturatedsodium bicarbonate, brine and dried over Na₂SO₄. The residue afterfiltration and evaporation of the solvent was chromatographed on silicagel eluting with 0.5:1:1 EtOAc:CH₂Cl₂:Hex to give the title compound asa white solid (0.94 g, 80%). mp 124-126° C. ¹H NMR (300 MHz, CDCl₃) δ6.00-6.10 (bs, 1H), 4.30-4.50 (m, 2H), 3.80-3.84 (m, 1H), 1.69 (s, 1H),1.36 (s, 3H), 1.32 (s, 3H). ¹³C NMR (75 MHz, CDCl₃) δ 160.5, 67.3, 62.9,46.4, 27.5, 27.4. LRMS (EI) m/z 162 (MH⁺), 179 (MNH₄ ⁺). Anal. Calcd forC₆H₁₁NO₂S.1/6 EtOAc: C, 45.52; H, 7.07; N, 7.96. Found: C, 45.83; H,6.86; N, 8.19.

[0205] 2e. 4-(1-Methyl-1-(nitrosothio)ethyl)-1,3-oxazolidin-2-one

[0206] To a solution of tert-butyl nitrite (1.7 mL of 90% solution, 1.48g, 14.4 mmol) in CH₂Cl₂ (2 mL) was added dropwise a solution of Example2d (0.94 g, 5.8 mmol) in CH₂Cl₂ (13 mL) at 0° C. The resulting greensolution was stirred at 0° C. for 20 minutes and then at roomtemperature for 15 minutes in the dark. The residue after evaporation ofthe solvent was chromatographed on silica gel eluting with 1:4EtOAc:CH₂Cl₂ to give the title compound as a purple-green solid (0.89 g,80%). mp 65° C. ¹H NMR (300 MHz, CDCl₃) δ 7.42 (bs, 1H), 4.40-4.65 (m,3H), 1.94 (s, 3H), 1.92 (s, 3H). ¹³C NMR (75 MHz, CDCl₃) δ 160.5, 67.0,61.3, 58.1, 25.3, 24.0. LRMS (EI) m/z 191 (MH⁺), 208 (MNH₄+). Anal.Calcd for C₆H₁₀N₂O₃S: C, 37.89; H, 5.30; N, 14.73; S, 16.85. Found: C,37.97; H, 5.26; N, 14.45; S, 16.78.

Example 3 Infrared Thermographic Measurements

[0207] Female white New Zealand rabbits were anaesthetized withpentobarbitol sodium and placed in a supine position on a warming pad.The warming pad was connected to a temperature control unit to maintainthe core (rectal) temperature to 38° C. The labia and clitoris wereexposed and kept in position by taping the surrounding skin to thenearby abdominal area. The infrared camera (THERMACAM® SC 1000,Inframetrics Inc., North Billerica, Mass.) was focused on the labia andclitoris and the animal was covered with a chamber to maintain the heatloss due to air movement.

[0208] After a steady baseline temperature was maintained and recordedfor at least 10 minutes the compound (50 μL) was applied to the surfaceof the labia and clitoris using a syringe and 27G needle. The compoundwas formulated in a mixture of dimethyl sulfoxide (25%) andpoly(ethylene glycols) (75%). The poly(ethylene glycols) was a mixtureof poly(ethylene glycol) 1450 and poly(ethylene glycol) 400 in a ratioof 1:9 respectively.

[0209] The images from the infrared camera were electronicallytransferred to a PC computer and analyzed using TherMonitor 95, version1.61 (Thermoteknix System Ltd., Mount pleasant, Cambridge, U. K.).Various color scales in the visible wave length are used to depict thetemperature change of the recorded images.

Example 4 Infrared Thermography Measurements Following TopicalAdministration of Isobutyl Nitrite

[0210] Infrared thermographic measurements were recorded as described inExample 3. FIGS. 1 and 2 shows the temperature increase of the rabbitgenitalia following the topical administration of 10% isobutyl nitriteor vehicle and 20% isobutyl nitrite or vehicle respectively. As can beseen from FIG. 1 (top panels), the temperature of the labia and clitorischanges from ˜30° C. prior to the application of the compound to 32° C.following the application of 10% isobutyl nitrite. As can be seen fromFIG. 2, application of 20% isobutyl nitrite results in a temperaturechange from ˜31° C. prior to the application of the compound to 34° C.following the application of 20% isobutyl nitrite. The application ofthe vehicle alone FIGS. 1 and 2 (bottom panels) did not result in atemperature increase.

Example 5 Infrared Thermography Measurements Following TopicalAdministration of Example 1

[0211] Infrared thermographic measurements were recorded as described inExample 3. FIG. 3 shows the temperature change of the rabbit genitaliafollowing the topical administration of Example 1 (5%) or vehicle. Ascan be seen from FIG. 3 (top panels), the temperature of the labia andclitoris changes from ˜32° C. prior to the application of the compoundto 35° C. following the application of Example 1. The application of thevehicle alone did not result in a temperature increase FIG. 3 (bottompanels).

Example 6 Infrared Thermography Measurements Following TopicalAdministration of Example 2

[0212] Infrared thermographic measurements were recorded as described inExample 3. FIG. 4 shows the temperature increase of the rabbit genitaliafollowing the topical administration of Example 2 (10%). As can be seenfrom FIG. 4 (top panel), the temperature of the labia and clitorischanges from ˜32° C. prior to the application of the compound to 36° C.following the application of Example 2. After 60 minutes thevasoconstrictor, phenylephrine (10%), was applied and the temperaturemonitored for an additional 5 minutes. As can be seen from FIG. 4(bottom panels), the addition of the vasoconstrictor resulted in adecrease in the temperature from 36° C. to 32° C. The results show thatthe temperature changes can be used as a measure of vasodilation of thetissue.

Example 7 Preparation of S-nitrosoglutathione

[0213] Glutathione (N-(N-L-,γ-glutamyl-L-cysteinyl)glycine) (100 g,0.325 mol) was dissolved in deoxygenated water (200 ml) and 2N HCl (162ml) at room temperature and then the reaction mixture was cooled to 0°C. With rapid stirring, a solution of sodium nitrite (24.4 g, 0.35 mol)in water (40 ml) was added. Stirring with cooling of the reactionmixture was continued for approximately 1 hour, after which time thepink precipitate which formed was collected by vacuum filtration. Thefilter cake was resuspended in chilled 40% acetone-water (600 ml) andcollected by vacuum filtration. The filter cake was washed with acetone(2×200 ml) and ether (100 ml) and then dried under high vacuum at roomtemperature in the dark to afford the title compound,N-(N-L-γ-glutamyl-S-Nitroso-L-cysteinyl)glycine, as a pink powder. ¹HNMR (D₂O): δ1.98 (m, 2H), 2.32 (t, 2H), 3.67 (t, 1H), 3.82 (s, 2H), 3.86(dd, 1H), 3.98 (dd, 1H), 4.53 (m, 1H).

Example 8 Infrared Thermography Measurements Following TopicalAdministration of Example 7

[0214] Infrared thermographic measurements were recorded as described inExample 3. FIG. 5 shows the temperature change of the rabbit genitaliafollowing topical application of Example 7 (50 μL, 2.5%), Example 7 (50μL, 5%), Example 7 (50 μL, 10%) or vehicle (50 μL, MIGLYOL®). As can beseen from FIG. 5, the temperature of the labia and clitoris increased by5.5° C. after application of 10% Example 7, and by 3.5° C. afterapplication of 5% Example 7. The application of 2.5% Example 7, orvehicle alone resulted in slight and transient increase of labialclitoral temperature.

Example 9 Laser Doppler Measurements of Labial/Clitoral Blood FlowFollowing Topical Administration of Example 7

[0215] Female white New Zealand rabbits were anaesthetized withKetamine/Xylazine mixture and placed in supine position. A laser dopplerprobe (LASERFLO, BPM 403, Vasamedics, Inc., MN) was placed at the labiumsurface, and after steady baseline blood flow was maintained andrecorded for at least 10 minutes, Example 5 (5%, 50, μL), 10% of Example7 (10%, 50 μL), or vehicle alone (50 μL, MIGLYOL®(D) was applied to thesurface of the labia and clitoris. As seen from FIG. 6, application ofExample 7, but not vehicle elicited a sustained (>60 min) increase(4-fold) in labial blood flow as compared to the baseline. The timecourse for the increase in blood flow (FIG. 6) was very similar to thetime course for the increase in labial clitoral temperature (FIG. 5).Hence the measured temperature increase could be the result of theincreased blood flow.

[0216] The disclosure of each patent, patent application and publicationcited or described in the specification is hereby incorporated byreference herein in its entirety.

[0217] Although the invention has been set forth in detail, one skilledin the art will appreciate that numerous changes and modifications canbe made to the invention without departing from the spirit and scopethereof.

What is claimed is:
 1. A compound selected from the group consisting of4-aza-4-(2-methyl-2-(nitrosothio)propyl)tricyclo(5.2.1.0<2,6>)dec-8-ene-3,5-dioneor a pharmaceutically acceptable salt thereof; and4-(1-methyl-1-(nitrosothio)ethyl)-1,3-oxazolidin-2-one or apharmaceutically acceptable salt thereof.
 2. A composition comprisingthe compound of claim 1 and a pharmaceutically acceptable carrier. 3.The composition of claim 3, further comprising at least one penetrationenhancer, at least one compound that donates, transfers or releasesnitric oxide, elevates endogenous levels of endothelium-derived relaxingfactor, stimulates endogenous synthesis of nitric oxide or is asubstrate for nitric oxide synthase, or a pharmaceutically acceptablesalt thereof and/or at least one vasoactive agent or a pharmaceuticallyacceptable salt thereof.
 4. The composition of claim 3, wherein the atleast one compound that donates, transfers or releases nitric oxide,elevates endogenous levels of endothelium-derived relaxing factor,stimulates endogenous synthesis of nitric oxide or is a substrate fornitric oxide synthase is an S-nitrosothiol.
 5. The composition of claim4, wherein the S-nitrosothiol is S-nitroso-N-acetylcysteine,S-nitroso-captopril, S-nitroso-N-acetylpenicillamine,S-nitroso-homocysteine, S-nitroso-cysteine, S-nitroso-glutathione orS-nitroso-cysteinyl-glycine.
 6. The composition of claim 5, wherein theS-nitrosothiol is S-nitroso-glutathione.
 7. The composition of claim 4,wherein the S-nitrosothiol is: (i) HS(C(R_(e))(R_(f)))_(m)SNO; (ii)ONS(C(R_(e))(R_(f)))_(m)R_(e); and (iii)H₂N—CH(CO₂H)—(CH₂)_(m)—C(O)NH—CH(CH₂SNO)—C(O)NH—CH₂—CO₂H; wherein m isan integer from 2 to 20; R_(e) and R_(f) are each independently ahydrogen, an alkyl, a cycloalkoxy, a halogen, a hydroxy, anhydroxyalkyl, an alkoxyalkyl, an arylheterocyclic ring, an alkylaryl, acycloalkylalkyl, a heterocyclicalkyl, an alkoxy, a haloalkoxy, an amino,an alkylamino, a dialkylamino, an arylamino, a diarylamino, analkylarylamino, an alkoxyhaloalkyl, a haloalkoxy, a sulfonic acid, asulfonic ester, an alkylsulfonic acid, an arylsulfonic acid, anarylalkoxy, an alkylthio, an arylthio, a cycloalkylthio, a cycloalkenyl,a cyano, an aminoalkyl, an aminoaryl, an aryl, an arylalkyl, analkylaryl, a carboxamido, a alkylcarboxamido, an arylcarboxamido, anamidyl, a carboxyl, a carbamoyl, a carbamate, an alkylcarboxylic acid,an arylcarboxylic acid, an alkylcarbonyl, an arylcarbonyl, an ester, acarboxylic ester, an alkylcarboxylic ester, an arylcarboxylic ester, ahaloalkoxy, a sulfonamido, an alkylsulfonamido, an arylsulfonamido, asulfonic ester, a urea, a phosphoryl, a nitro, -T-Q, or—(C(R_(e))(R_(f)))_(k)-T-Q, or R_(e) and R_(f) taken together with thecarbon atom to which they are attached form a carbonyl, a methanthial, aheterocyclic ring, a cycloalkyl group or a bridged cycloalkyl group; Qis —NO or —NO₂; and T is independently a covalent bond, a carbonyl, anoxygen, —S(O)_(o)— or —N(R_(a))R_(i)—, wherein o is an integer from 0 to2, R_(a) is a lone pair of electrons, a hydrogen or an alkyl group;R_(i) is a hydrogen, an alkyl, an aryl, an alkylcarboxylic acid, an arylcarboxylic acid, an alkylcarboxylic ester, an arylcarboxylic ester, analkylcarboxamido, an arylcarboxamido, an alkylaryl, an alkylsulfinyl, analkylsulfonyl, an arylsulfinyl, an arylsulfonyl, a sulfonamido, acarboxamido, a carboxylic ester, an amino alkyl, an amino aryl,—CH₂—C(T-Q)(R_(e))(R_(f)), or —(N₂O₂—).M⁺, wherein M⁺ is an organic orinorganic cation; with the proviso that when R_(i) is—CH₂—C(T-Q)(R_(e))(R_(f)) or —(N₂O₂—).M⁺; then “-T-Q” can be a hydrogen,an alkyl group, an alkoxyalkyl group, an aminoalkyl group, a hydroxygroup or an aryl group.
 8. The composition of claim 3, wherein the atleast one compound that donates, transfers or releases nitric oxide,elevates endogenous levels of endothelium-derived relaxing factor,stimulates endogenous synthesis of nitric oxide or is a substrate fornitric oxide synthase is: (i) a compound that comprises at least oneON—O—, ON—N— or ON—C— group; (ii) a compound that comprises at least oneO₂N—O—, O₂N—N—, O₂N—S— or —O₂N—C— group; (iii) a N-oxo-N-nitrosoaminehaving the formula: R¹R²N—N(O-M⁺)—NO, wherein R¹ and R² are eachindependently a polypeptide, an amino acid, a sugar, an oligonucleotide,a straight or branched, saturated or unsaturated, aliphatic or aromatic,substituted or unsubstituted hydrocarbon, or a heterocyclic group, andM⁺is an organic or inorganic cation.
 9. The composition of claim 8,wherein the compound comprising at least one ON—O—, ON—N— or ON—C— groupis an ON—O-polypeptide, an ON—N-polypepetide, an ON—C-polypeptide, anON—O-amino acid, an ON—N-amino acid, an ON—C-amino acid, an ON—O-sugar,an ON—N-sugar, an ON—C-sugar, an ON—O-oligonucleotide, anON—N-oligonucleotide, an ON—C-oligonucleotide, a straight or branched,saturated or unsaturated, substituted or unsubstituted, aliphatic oraromatic ON—O-hydrocarbon, a straight or branched, saturated orunsaturated, substituted or unsubstituted, aliphatic or aromaticON—N-hydrocarbon, a straight or branched, saturated or unsaturated,substituted or unsubstituted, aliphatic or aromatic ON—C-hydrocarbon, anON—O-heterocyclic compound, an ON—N-heterocyclic compound or aON—C-heterocyclic compound.
 10. The composition of claim 8, whereincompound comprising at least one O₂N—O—, O₂N—N—, O₂N—S— or O₂N—C— groupis an O₂N—O-polypeptide, an O₂N—N-polypeptide, an O₂N—S-polypeptide, anO₂N—C-polypeptide, an O₂N—O-amino acid, O₂N—N-amino acid, O₂N—S-aminoacid, an O₂N—C-amino acid, an O₂N—O-sugar, an O₂N—N-sugar, O₂N—S-sugar,an O₂N—C-sugar, an O₂N—O-oligonucleotide, an O₂N—N-oligonucleotide, anO₂N—S oligonucleotide, an O₂N—C-oligonucleotide, a straight or branched,saturated or unsaturated, aliphatic or aromatic, substituted orunsubstituted O₂N—O-hydrocarbon, a straight or branched, saturated orunsaturated, aliphatic or aromatic, substituted or unsubstitutedO₂N—N-hydrocarbon, a straight or branched, saturated or unsaturated,aliphatic or aromatic, substituted or unsubstituted O₂N—S-hydrocarbon, astraight or branched, saturated or unsaturated, aliphatic or aromatic,substituted or unsubstituted O₂N—C-hydrocarbon, an O₂N—O-heterocycliccompound, an O₂N—N-heterocyclic compound, an O₂N—S-heterocyclic compoundor an O₂N—C-heterocyclic compound.
 11. The composition of claim 3,wherein the at least one compound that donates, transfers or releasesnitric oxide, elevates endogenous levels of endothelium-derived relaxingfactor, stimulates endogenous synthesis of nitric oxide or is asubstrate for nitric oxide synthase is L-arginine, L-homoarginine,N-hydroxy-L-arginine, nitrosated L-arginine, nitrosylated L-arginine,nitrosated N-hydroxy-L-arginine, nitrosylated N-hydroxy-L-arginine,citrulline, ornithine, glutamine, lysine, polypeptides comprising atleast one of these amino acids or inhibitors of the enzyme arginase. 12.The composition of claim 3, wherein the at least one compound thatdonates, transfers or releases nitric oxide, elevates endogenous levelsof endothelium-derived relaxing factor, stimulates endogenous synthesisof nitric oxide or is a substrate for nitric oxide synthase is aNONOate.
 13. The composition of claim 3, wherein the vasoactive agent isa potassium channel activator, a calcium channel blocker, anα-adrenergic receptor antagonist, a β-blocker, a phosphodiesteraseinhibitor, adenosine, an ergot alkaloid, a vasoactive intestinalpeptide, a prostaglandin, a dopamine agonist, an opioid antagonist, anendothelin antagonist, a thromboxane inhibitor or a mixture thereof. 14.The composition of claim 3, wherein the penetration enhancer isdimethylsulfoxide, dimethyl formamide, N,N-dimethylacetamide,decylmethylsulfoxide, polyethylene glycol monolaurate,polyethyleneglycol, glycerol monolaurate, lecithin, a 1-substitutedazacycloheptan-2-one, a lower alkanol, a C₆ to C₂₀-hydrocarbylsubstituted 1,3-dioxane, a C₆ to C₂₀-hydrocarbyl substituted1,3-dioxolane or a C₆ to C₂₀-hydrocarbyl substituted acetal, analkonate, a glyceride, a surfactant, or a mixture thereof.
 15. Thecomposition of claim 14, wherein the glyceride is a mono glyceride, adiglyceride, a triglycerides, a polyglycolyzed glyceride or a mixturethereof.
 16. The composition of claim 15, wherein the glyceride is amixture of caprylic triglycerides and capric triglycerides, a decanolytriglyceride, an octanoyl triglyceride, a C₈-C₁₂ triglyceride, asaturated polyglycolyzed glyceride, a glyceryl caprylate/caprate andPEG-8 (polyethylene glycol) caprylate/caprate complex, a unsaturatedpolyglycolyzed glyceride, an apricot kernel oil PEG-6 complex, an almondoil PEG-6 complex, a peanut oil PEG-6 complex, an olive oil PEG-6complex, a corn oil PEG-6 complex, an ethoxylated glyceride, a glycerylcaprylate/caprate PEG-4 complex, or a mixture thereof.
 17. A method fortreating a sexual dysfunction in a patient in need thereof comprisingadministering to the patient a therapeutically effective amount of thecomposition of claim
 2. 18. The method of claim 17, wherein the patientis female.
 19. The method of claim 17, wherein the patient is male. 20.The method of claim 17, wherein the composition is administered orally,bucally, topically, by injection, by inhalation or by transurethralapplication.
 21. The method of claim 20, wherein the composition isadministered orally as a solid or liquid dose.
 22. The method of claim17, further comprising administering at least one compound that donates,transfers or releases nitric oxide, elevates endogenous levels ofendothelium-derived relaxing factor, stimulates endogenous synthesis ofnitric oxide or is a substrate for nitric oxide synthase and/or the atleast one vasoactive agent
 23. The method of claim 22, wherein the atleast one compound that donates, transfers or releases nitric oxide,elevates endogenous levels of endothelium-derived relaxing factor,stimulates endogenous synthesis of nitric oxide or is a substrate fornitric oxide synthase and the at least one vasoactive agent areadministered separately.
 24. The method of claim 22, wherein the atleast one compound that donates, transfers or releases nitric oxide,elevates endogenous levels of endothelium-derived relaxing factor,stimulates endogenous synthesis of nitric oxide or is a substrate fornitric oxide synthase and the at least one vasoactive agent areadministered in the form of a composition.
 25. A kit comprising at leastone compound of claim
 1. 26. The kit of claim 25, further comprising atleast one penetration enhancer, at least one compound that donates,transfers or releases nitric oxide, elevates endogenous levels ofendothelium-derived relaxing factor, stimulates endogenous synthesis ofnitric oxide or is a substrate for nitric oxide synthase and/or at leastone vasoactive agent.
 27. A compound selected from the group consistingof4-aza4-(2-methyl-2-sulfanylpropyl)tricyclo(5.2.1.0<2,6>)dec-8-ene-3,5-dioneor a pharmaceutically acceptable salt thereof;4-1{1-methyl-1-((2,4,6-trimethoxyphenyl)methylthio)ethyl}-1,3-oxazolidin-2-oneor a pharmaceutically acceptable salt thereof; and2-amino-3-methyl-3-((2,4,6-trimethoxyphenyl)methylthio)butan-1-ol or apharmaceutically acceptable salt thereof. 28.4-aza-4-(2-methyl-2-(nitrosothio)propyl)tricyclo(5.2.1.0<2,6>)dec-8-ene-3,5-dioneor a pharmaceutically acceptable salt thereof. 29.4-(1-methyl-1-(nitrosothio)ethyl)-1,3-oxazolidin-2-one or apharmaceutically acceptable salt thereof.